TITLE OF THE INVENTION Human binding molecules capable of neutralizing influenza A viruses of phylogenetic group 1 and phylogenetic group 2 and influenza B viruses FIELD OF THE INVENTION The invention relates to medicine. The invention in particular relates to human binding molecules capable of neutralizing influenza A viruses of both phylogenetic group 1 and phylogenetic group 2. In particular, the invention relates to binding molecules capable of neutralizing influenza A viruses of both phylogenetic group 1 and phylogenetic group 2, as well as influenza B viruses. The invention further relates to the diagnosis, prophylaxis and/or treatment of an infection caused by influenza A viruses of phylogenetic groups 1 and 2, and preferably also influenza B viruses.

BACKGROUND OF THE INVENTION Influenza infection (also referred to as “influenza” or “the flu”) is one of the most common diseases known to man causing between three and five million cases of severe illness and between 250,000 and 500,000 deaths every year around the world. Influenza rapidly spreads in seasonal epidemics affecting 5-15% of the population and the burden on health care costs and lost productivity are extensive (World Healthcare Organization (WHO)).

There are 3 types of influenza virus (types A, B and C) responsible for infectious pathologies in humans and animals. The type A and type B viruses are the agents responsible for the influenza seasonal epidemics and pandemics observed in humans.

Influenza A viruses can be classified into influenza virus subtypes based on variations in antigenic regions of two genes that encode the surface glycoproteins hemagglutinin (HA) and neuraminidase (NA) which are required for viral attachment and cellular release. Currently, sixteen subtypes of HA (H1-H16) and nine NA (N1-N9) antigenic variants are known in influenza A virus. Influenza virus subtypes can further be classified by reference to their phylogenetic group. Phylogenetic analysis (Fouchier et al., 2005) has demonstrated a subdivision of HAs comprising two main groups (Air, 1981): inter alia the H1, H2, H5 and H9 subtypes in phylogenetic group 1 (herein also referred to as “group 1”) and inter alia the H3, H4 and H7 subtypes in phylogenetic group 2 (or “group 2”). Only some of the influenza A subtypes (i.e. H1N1, H1N2 and H3N2) circulate among people, but all combinations of the 16 HA and 9 NA subtypes have been identified in animals, in particular in avian species. Animals infected with influenza A often act as a reservoir for the influenza viruses and certain subtypes have been shown to cross the species barrier to humans, such as the highly pathogenic influenza A strain H5N1.

The influenza type B virus strains are strictly human. The antigenic variations in HA within the influenza type B virus strains are weaker than those observed within the type A strains. Two genetically and antigenically distinct lineages of influenza B virus are circulating in humans, as represented by the B/Yamagata/16/88 (also referred to as B/Yamagata) and B/Victoria/2/87 (B/Victoria) lineages (Ferguson et al., 2003). Although the spectrum of disease caused by influenza B viruses is generally milder than that caused by influenza A viruses, severe illness requiring hospitalization is still frequently observed with influenza B infection.

Current approaches to dealing with annual influenza epidemics include annual vaccination, preferably generating heterotypic cross-protection. However, circulating influenza viruses in humans are subject to permanent antigenic changes which require annual adaptation of the influenza vaccine formulation to ensure the closest possible match between the influenza vaccine strains and the circulating influenza strains.

Although yearly vaccination with influenza vaccines is the best way to prevent influenza, antiviral drugs, such as oseltamivir (Tamiflu®) can be effective for prevention and treatment of influenza infection. The number of influenza virus strains showing resistance against antiviral drugs, such as oseltamivir is, however, increasing.

An alternative approach is the development of antibody-based prophylactic or therapeutic treatments to neutralize various seasonal and pandemic influenza viruses. The primary target of most neutralizing antibodies that protect against influenza virus infection is the globular head (HA1 part) of the viral HA protein which contains the receptor binding site, but which is subject to continuing genetic evolution with amino acid substitutions in antibody-binding sites (antigenic drift).

Recently, broadly cross-neutralizing antibodies recognizing an epitope in the conserved stem region of hemagglutinin of influenza A viruses of phylogenetic group 1 (including e.g. the H1 and H5 influenza subtypes) have been identified (see e.g.

WO2008/028946), as well as cross-neutralizing antibodies recognizing a highly conserved epitope in the stem region of HA of influenza A viruses of phylogenetic group 2 (including e.g. H3 and H7 subtypes) (). The neutralizing activity of these antibodies is restricted to either group 1 or group 2 influenza viruses. In addition, these antibodies are not capable of binding to and neutralizing influenza B viruses.

Furthermore, discloses a human antibody FI6 binding to hemagglutinin and capable of binding to and neutralizing influenza A subtypes of group 1 (including H1 and H5 subtypes) and group 2 (including H3 and H7 subtypes). This antibody also does not bind HA from influenza B viruses.

In addition, US 2009/0092620 discloses a murine antibody recognizing an antigenic structure present in hemagglutinin of both the H1 and the H3 subtype and on hemagglutinin of influenza B viruses belonging to the B/Victoria and B/Yamagata groups. The antibodies inhibit the hemagglutination activity of several H3N2 strains implicating that this antibody binds an epitope in the globular head of HA.

In view of the severity of the respiratory illness caused by influenza A and influenza B viruses, as well has the high economic impact of the seasonal epidemics, and the continuing risk for pandemics, there is an ongoing need for effective means for the prevention and treatment of influenza A and B subtypes. There is thus a need for binding molecules, preferably broadly neutralizing human binding molecules, capable of cross- neutralizing influenza A viruses of both phylogenetic group 1 and phylogenetic group 2, and preferably also influenza B viruses. It is an object of the present invention to go some way towards meeting this need and/or to provide the public with a useful choice.

SUMMARY OF THE INVENTION In a first aspect, the invention provides an isolated binding molecule, capable of specifically binding to an epitope in the stem region of the hemagglutinin protein (HA) of influenza A virus subtypes of phylogenetic group 1 and influenza A virus subtypes of phylogenetic group 2 subtypes, and capable of neutralizing influenza A virus subtypes of phylogenetic group 1 and phylogenetic group 2, characterized in that the binding molecule is also capable of specifically binding to the hemagglutinin protein (HA) of influenza B virus subtypes, wherein the binding molecule is selected from the group consisting of: a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, and a light chain CDR1 region of SEQ ID NO: 146, a light chain CDR2 region of SEQ ID NO: 174, and a light chain CDR3 region of SEQ ID NO: 147, a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, and a light chain CDR1 region of SEQ ID NO: 148, a light chain CDR2 region of SEQ ID NO: 149, and a light chain CDR3 region of SEQ ID NO: 150, a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, and a light chain CDR1 region of SEQ ID NO: 142, a light chain CDR2 region of SEQ ID NO: 143, and a light chain CDR3 region of SEQ ID NO: 173; and a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, and a light chain CDR1 region of SEQ ID NO: 148, a light chain CDR2 region of SEQ ID NO: 149, and a light chain CDR3 region of SEQ ID NO: 150, a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, and a light chain CDR1 region of SEQ ID NO: 156, a light chain CDR2 region of SEQ ID NO: 157, and a light chain CDR3 region of SEQ ID NO: 158, a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, and a light chain CDR1 region of SEQ ID NO: 171, a light chain CDR2 region of SEQ ID NO: 164, and a light chain CDR3 region of SEQ ID NO: 172, and a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, and a light chain CDR1 region of SEQ ID NO: 142, a light chain CDR2 region of SEQ ID NO: 143, and a light chain CDR3 region of SEQ ID NO: 144.

In a second aspect, the invention provides a nucleic acid molecule encoding a binding molecule according to the invention.

In a third aspect, the invention provides a pharmaceutical composition comprising a binding molecule according to the invention, and/or a nucleic acid molecule according to the invention, and a pharmaceutically acceptable excipient.

In a fourth aspect, the invention provides a use of a binding molecule of the invention, and/or a nucleic acid molecule of the invention, in the manufacture of a medicament.

In a fifth aspect, the invention provides a use of a binding molecule of the invention, and/or a nucleic acid molecule of the invention, in the manufacture of a medicament for the diagnostic, therapeutic and/or prophylactic treatment of influenza infection.

Also described are binding molecules capable of specifically binding to influenza A virus strains from both phylogenetic group 1 (including e.g. influenza viruses comprising HA of the H1 and H5 subtype) and influenza A virus strains from phylogenetic group 2 (including e.g. influenza viruses comprising HA of the H3 and H7 subtype). In an embodiment, the binding molecules also have neutralizing activity against influenza A virus strains from both phylogenetic group 1 and phylogenetic group 2. In an embodiment, the binding molecules are furthermore capable of specifically binding influenza B virus strains, including e.g. influenza B virus strains of the B/Yamagata and/or B/Victoria lineages. In an embodiment, the binding molecules are furthermore capable of neutralizing influenza B virus strains, including e.g. influenza B virus strains of the B/Yamagata and/or B/Victoria lineages. In an embodiment, the binding molecules are capable of in vivo neutralizing influenza A and/or B virus strains. In an embodiment the binding molecules bind to a conserved epitope in the stem region of the HA protein of influenza A and B viruses. In an embodiment, the binding molecules have no hemagglutination inhibiting (HI) activity.

Also described are binding molecules that bind to an epitope in the stem region of the haemagglutinin protein that is shared between influenza A virus subtypes within the phylogenetic group 1 and influenza virus subtypes within phylogenetic group 2, as well as influenza B virus subtypes, and therefore relates to binding molecules that cross-react between both group 1 and group 2 influenza A virus subtypes and influenza B viruses.

The invention also pertains to nucleic acid molecules encoding at least the binding region of the human binding molecules.

The binding molecules and/or nucleic acid molecules described herein are suitable for use as a universal prophylactic, diagnostic and/or treatment agent for influenza A viruses and influenza B viruses, even irrespective of the causative influenza subtype.

It is surmised that the binding molecules described herein bind to hitherto unknown and highly conserved epitopes that are not or much less prone to antigenic drift or shift. In particular, this epitope is shared between influenza viruses belonging to both phylogenetic group 1 and phylogenetic group 2, and influenza B viruses. It is also encompassed to use the binding molecules of the invention to identify and/or characterize these epitopes.

Also described is the use of the human binding molecules and/or the nucleic acid molecules described herein in the diagnosis, prophylaxis and/or treatment of a subject having, or at risk of developing, an influenza virus infection. Also described is the use of the human binding molecules and/or the nucleic acid molecules of the invention in the diagnosis/detection of such influenza infections.

DESCRIPTION OF THE FIGURES shows the blocking of conformational change of H1, H5, H9, H3, and H7 HAs by CR9114. (A) FACS binding of CR9114 to various conformations – uncleaved precursor (HA0); neutral pH, cleaved (HA); fusion pH, cleaved (fusion pH) – of surface-expressed rHA of A/New Caledonia/20/1999 (H1) A/Viet Nam/1203/2004 (H5), A/Hong Kong/1073/1999 (H9), A/Wisconsin/67/2005 (H3), and A/Netherlands/219/2003 (H7).

Binding is expressed as the percentage of binding to untreated rHA (HA0). (B) FACS binding of CR9114 to surface-expressed HA as above, except that mAb CR9114 was added before exposure of the cleaved HAs to a pH of 4.9. shows that MAb CR9114 competes with CR6261 and CR8020 for binding to H1 and H3, respectively. Additional degree of binding of indicated mAbs to immobilized HA of A/New Caledonia/20/1999 (H1N1) saturated with 100 nM of CR6261 or CR9114 (panels A and B), or to immobilized HA of A/Wisconsin/67/2005 (H3N2) saturated with 100 nM of CR8020 or CR9114 (panels C and D), measured using biolayer interferometry. demonstrates the prophylactic efficacy of CR9114 in the mouse lethal challenge model with influenza B (B/Florida/04/2006) virus. A. Kaplan-Meier survival curves of mice treated intravenously with either 15 mg/kg CR9114 or vehicle control on day -1 before challenge, followed by a challenge at day 0 of 25 LD B/Florida/04/2006. B. Mean bodyweight change (%) relative to day 0. Bars represent 95% CI of the mean. If a mouse died/was euthanized during the study, the last observed bodyweight was carried forward.

C. Median Clinical scores. Bars represent interquartile ranges. Clinical score explanation: 0=no clinical signs; 1=rough coat; 2=rough coat, less reactive during handling; 3=rough coat, rolled up, laboured breathing, less reactive during handling; 4=rough coat, rolled up, laboured breathing, inactive response to manipulation/handlings.

DESCRIPTION OF THE INVENTION Definitions of terms as used in the present invention are given below.

The term “comprising” as used in this specification and claims means “consisting at least in part of”. When interpreting statements in this specification, and claims which include the term “comprising”, it is to be understood that other features that are additional to the features prefaced by this term in each statement or claim may also be present.

Related terms such as “comprise” and “comprised” are to be interpreted in similar manner.

The term “included” or “including” as used herein is deemed to be followed by the words “without limitation”.

As used herein the term “binding molecule” refers to an intact immunoglobulin including monoclonal antibodies, such as chimeric, humanized or human monoclonal antibodies, or to an antigen-binding and/or variable domain comprising fragment of an immunoglobulin that competes with the intact immunoglobulin for specific binding to the binding partner of the immunoglobulin, e.g. HA. Regardless of structure, the antigen- binding fragment binds with the same antigen that is recognized by the intact immunoglobulin. An antigen-binding fragment can comprise a peptide or polypeptide comprising an amino acid sequence of at least 2, 5, 10, 15, 20, 25, 30, 35, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, 200, or 250 contiguous amino acid residues of the amino acid sequence of the binding molecule.

The term “binding molecule”, as used herein includes all immunoglobulin classes and subclasses known in the art. Depending on the amino acid sequence of the constant domain of their heavy chains, binding molecules can be divided into the five major classes of intact antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgA1, IgA2, IgG1, IgG2, IgG3 and IgG4.

Antigen-binding fragments include, inter alia, Fab, F(ab'), F(ab')2, Fv, dAb, Fd, complementarity determining region (CDR) fragments, single-chain antibodies (scFv), bivalent single-chain antibodies, single-chain phage antibodies, diabodies, triabodies, tetrabodies, (poly)peptides that contain at least a fragment of an immunoglobulin that is sufficient to confer specific antigen binding to the (poly)peptide, etc. The above fragments may be produced synthetically or by enzymatic or chemical cleavage of intact immunoglobulins or they may be genetically engineered by recombinant DNA techniques. The methods of production are well known in the art and are described, for example, in Antibodies: A Laboratory Manual, Edited by: E. Harlow and D, Lane (1988), Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, which is incorporated herein by reference. A binding molecule or antigen-binding fragment thereof may have one or more binding sites. If there is more than one binding site, the binding sites may be identical to one another or they may be different.

The binding molecule can be a naked or unconjugated binding molecule but can also be part of an immunoconjugate. A naked or unconjugated binding molecule is intended to refer to a binding molecule that is not conjugated, operatively linked or otherwise physically or functionally associated with an effector moiety or tag, such as inter alia a toxic substance, a radioactive substance, a liposome, an enzyme. It will be understood that naked or unconjugated binding molecules do not exclude binding molecules that have been stabilized, multimerized, humanized or in any other way manipulated, other than by the attachment of an effector moiety or tag. Accordingly, all post-translationally modified naked and unconjugated binding molecules are included herewith, including where the modifications are made in the natural binding molecule- producing cell environment, by a recombinant binding molecule-producing cell, and are introduced by the hand of man after initial binding molecule preparation. Of course, the term naked or unconjugated binding molecule does not exclude the ability of the binding molecule to form functional associations with effector cells and/or molecules after administration to the body, as some of such interactions are necessary in order to exert a biological effect. The lack of associated effector group or tag is therefore applied in definition to the naked or unconjugated binding molecule in vitro, not in vivo.

As used herein, the term “biological sample” encompasses a variety of sample types, including blood and other liquid samples of biological origin, solid tissue samples such as a biopsy specimen or tissue cultures, or cells derived there from and the progeny thereof. The term also includes samples that have been manipulated in any way after their procurement, such as by treatment with reagents, solubilization, or enrichment for certain components, such as proteins or polynucleotides. The term encompasses various kinds of clinical samples obtained from any species, and also includes cells in culture, cell supernatants and cell lysates.

The term “complementarity determining regions” (CDR) as used herein means sequences within the variable regions of binding molecules, such as immunoglobulins, that usually contribute to a large extent to the antigen binding site which is complementary in shape and charge distribution to the epitope recognized on the antigen.

The CDR regions can be specific for linear epitopes, discontinuous epitopes, or conformational epitopes of proteins or protein fragments, either as present on the protein in its native conformation or, in some cases, as present on the proteins as denatured, e.g., by solubilization in SDS. Epitopes may also consist of posttranslational modifications of proteins.

The term “deletion”, as used herein, denotes a change in either amino acid or nucleotide sequence in which one or more amino acid or nucleotide residues, respectively, are absent as compared to the reference, often the naturally occurring, molecule.

The term “expression-regulating nucleic acid sequence” as used herein refers to polynucleotide sequences necessary for and/or affecting the expression of an operably linked coding sequence in a particular host organism. The expression-regulating nucleic acid sequences, such as inter alia appropriate transcription initiation, termination, promoter, enhancer sequences; repressor or activator sequences; efficient RNA processing signals such as splicing and polyadenylation signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (e.g., ribosome binding sites); sequences that enhance protein stability; and when desired, sequences that enhance protein secretion, can be any nucleic acid sequence showing activity in the host organism of choice and can be derived from genes encoding proteins, which are either homologous or heterologous to the host organism. The identification and employment of expression-regulating sequences is routine to the person skilled in the art.

The term “functional variant”, as used herein, refers to a binding molecule that comprises a nucleotide and/or amino acid sequence that is altered by one or more nucleotides and/or amino acids compared to the nucleotide and/or amino acid sequences of the reference binding molecule and that is capable of competing for binding to the binding partner, i.e. the influenza virus, with the reference binding molecule. In other words, the modifications in the amino acid and/or nucleotide sequence of the reference binding molecule do not significantly affect or alter the binding characteristics of the binding molecule encoded by the nucleotide sequence or containing the amino acid sequence, i.e. the binding molecule is still able to recognize and bind its target. The functional variant may have conservative sequence modifications including nucleotide and amino acid substitutions, additions and deletions. These modifications can be introduced by standard techniques known in the art, such as site-directed mutagenesis and random PCR-mediated mutagenesis, and may comprise natural as well as non-natural nucleotides and amino acids.

Conservative amino acid substitutions include the ones in which the amino acid residue is replaced with an amino acid residue having similar structural or chemical properties. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chains (e.g., glycine, alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan). It will be clear to the skilled artisan that also other classifications of amino acid residue families than the one used above can be employed. Furthermore, a variant may have non-conservative amino acid substitutions, e.g., replacement of an amino acid with an amino acid residue having different structural or chemical properties. Similar minor variations may also include amino acid deletions or insertions, or both. Guidance in determining which amino acid residues may be substituted, inserted, or deleted without abolishing immunological activity may be found using computer programs well known in the art.

A mutation in a nucleotide sequence can be a single alteration made at a locus (a point mutation), such as transition or transversion mutations, or alternatively, multiple nucleotides may be inserted, deleted or changed at a single locus. In addition, one or more alterations may be made at any number of loci within a nucleotide sequence. The mutations may be performed by any suitable method known in the art.

The term “influenza virus subtype” as used herein in relation to influenza A viruses refers to influenza A virus variants that are characterized by various combinations of the hemagglutinin (H) and neuramidase (N) viral surface proteins. According to the present invention influenza virus subtypes may be referred to by their H number, such as for example “influenza virus comprising HA of the H1 or H3 subtype”, or “H1 influenza virus” “H3 influenza virus”, or by a combination of a H number and an N number, such as for example “influenza virus subtype H3N2” or “H3N2”.

The term influenza virus “subtype” specifically includes all individual influenza virus “strains” within each subtype, which usually result from mutations and show different pathogenic profiles. Such strains may also be referred to as various “isolates” of a viral subtype. Accordingly, as used herein, the terms “strains” and “isolates” may be used interchangeably. The current nomenclature for human influenza virus strains or isolates includes the geographical location of the first isolation, strain number and year of isolation, usually with the antigenic description of HA and NA given in brackets, e.g.

A/Moscow/10/00 (H3N2). Non-human strains also include the host of origin in the nomenclature.

The term “neutralizing” as used herein in relation to the binding molecules described herein refers to binding molecules that inhibit an influenza virus from replicatively infecting a target cell, regardless of the mechanism by which neutralization is achieved. Thus, neutralization can e.g. be achieved by inhibiting the attachment or adhesion of the virus to the cell surface, or by inhibition of the fusion of viral and cellular membranes following attachment of the virus to the target cell, and the like.

The term “cross-neutralizing” or “cross-neutralization” as used herein in relation to the binding molecules described herein refers to the ability of the binding molecules described herein to neutralize different subtypes of influenza A and/or B viruses.

The term “host”, as used herein, is intended to refer to an organism or a cell into which a vector such as a cloning vector or an expression vector has been introduced. The organism or cell can be prokaryotic or eukaryotic. Preferably, the hosts isolated host cells, e.g. host cells in culture. The term “host cells” merely signifies that the cells are modified for the (over)-expression of the binding molecules described herein and include B-cells that originally express these binding molecules and which cells have been modified to over-express the binding molecule by immortalization, amplification, enhancement of expression etc. It should be understood that the term host is intended to refer not only to the particular subject organism or cell but to the progeny of such an organism or cell as well. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent organism or cell, but are still included within the scope of the term “host” as used herein.

The term “human”, when applied to binding molecules as defined herein, refers to molecules that are either directly derived from a human or based upon a human germ line sequence. When a binding molecule is derived from or based on a human sequence and subsequently modified, it is still to be considered human as used throughout the specification. In other words, the term human, when applied to binding molecules is intended to include binding molecules having variable and constant regions derived from human germline immunoglobulin sequences or based on variable or constant regions occurring in a human or human lymphocyte and modified in some form. Thus, the human binding molecules may include amino acid residues not encoded by human germline immunoglobulin sequences, comprise substitutions and/or deletions (e.g., mutations introduced by for instance random or site-specific mutagenesis in vitro or by somatic mutation in vivo). “Based on” as used herein refers to the situation that a nucleic acid sequence may be exactly copied from a template, or with minor mutations, such as by error-prone PCR methods, or synthetically made matching the template exactly or with minor modifications.

The term “insertion”, also known as the term “addition”, denotes a change in an amino acid or nucleotide sequence resulting in the addition of one or more amino acid or nucleotide residues, respectively, as compared to the parent sequence.

The term “isolated”, when applied to binding molecules as defined herein, refers to binding molecules that are substantially free of other proteins or polypeptides, particularly free of other binding molecules having different antigenic specificities, and are also substantially free of other cellular material and/or chemicals. For example, when the binding molecules are recombinantly produced, they are preferably substantially free of culture medium components, and when the binding molecules are produced by chemical synthesis, they are preferably substantially free of chemical precursors or other chemicals, i.e., they are separated from chemical precursors or other chemicals which are involved in the synthesis of the protein. The term “isolated” when applied to nucleic acid molecules encoding binding molecules as defined herein, is intended to refer to nucleic acid molecules in which the nucleotide sequences encoding the binding molecules are free of other nucleotide sequences, particularly nucleotide sequences encoding binding molecules that bind other binding partners. Furthermore, the term “isolated” refers to nucleic acid molecules that are substantially separated from other cellular components that naturally accompany the native nucleic acid molecule in its natural host, e.g., ribosomes, polymerases, or genomic sequences with which it is naturally associated.

Moreover, “isolated” nucleic acid molecules, such as cDNA molecules, can be substantially free of other cellular material, or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized.

The term “monoclonal antibody” as used herein refers to a preparation of antibody molecules of single specificity. A monoclonal antibody displays a single binding specificity and affinity for a particular epitope. Accordingly, the term “human monoclonal antibody” refers to an antibody displaying a single binding specificity which has variable and constant regions derived from or based on human germline immunoglobulin sequences or derived from completely synthetic sequences. The method of preparing the monoclonal antibody is not relevant for the binding specificity.

The term “naturally occurring” as used herein as applied to an object refers to the fact that an object or compound can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism that can be isolated from a source in nature and which has not been intentionally modified by man in the laboratory is naturally occurring.

The term “nucleic acid molecule” as used herein refers to a polymeric form of nucleotides and includes both sense and anti-sense strands of RNA, cDNA, genomic DNA, and synthetic forms and mixed polymers of the above. A nucleotide refers to a ribonucleotide, deoxynucleotide or a modified form of either type of nucleotide. The term also includes single- and double-stranded forms of DNA. In addition, a polynucleotide may include either or both naturally occurring and modified nucleotides linked together by naturally occurring and/or non-naturally occurring nucleotide linkages. The nucleic acid molecules may be modified chemically or biochemically or may contain non-natural or derivatized nucleotide bases, as will be readily appreciated by those of skill in the art.

Such modifications include, for example, labels, methylation, substitution of one or more of the naturally occurring nucleotides with an analogue, internucleotide modifications such as uncharged linkages (e.g., methyl phosphonates, phosphotriesters, phosphoramidates, carbamates, etc.), charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), pendent moieties (e.g., polypeptides), intercalators (e.g., acridine, psoralen, etc.), chelators, alkylators, and modified linkages (e.g., alpha anomeric nucleic acids, etc.). The above term is also intended to include any topological conformation, including single-stranded, double-stranded, partially duplexed, triplex, hairpinned, circular and padlocked conformations. Also included are synthetic molecules that mimic polynucleotides in their ability to bind to a designated sequence via hydrogen bonding and other chemical interactions. Such molecules are known in the art and include, for example, those in which peptide linkages substitute for phosphate linkages in the backbone of the molecule. A reference to a nucleic acid sequence encompasses its complement unless otherwise specified. Thus, a reference to a nucleic acid molecule having a particular sequence should be understood to encompass its complementary strand, with its complementary sequence. The complementary strand is also useful, e.g., for anti-sense therapy, hybridisation probes and PCR primers.

The term “operably linked” refers to two or more nucleic acid sequence elements that are usually physically linked and are in a functional relationship with each other. For instance, a promoter is operably linked to a coding sequence, if the promoter is able to initiate or regulate the transcription or expression of a coding sequence, in which case the coding sequence should be understood as being “under the control of” the promoter.

By “pharmaceutically acceptable excipient” is meant any inert substance that is combined with an active molecule such as a drug, agent, or binding molecule for preparing an agreeable or convenient dosage form. The “pharmaceutically acceptable excipient” is an excipient that is non-toxic to recipients at the used dosages and concentrations, and is compatible with other ingredients of the formulation comprising the drug, agent or binding molecule. Pharmaceutically acceptable excipients are widely applied and known in the art.

The term “specifically binding”, as used herein, in reference to the interaction of a binding molecule, e.g. an antibody, and its binding partner, e.g. an antigen, means that the interaction is dependent upon the presence of a particular structure, e.g. an antigenic determinant or epitope, on the binding partner. In other words, the antibody preferentially binds or recognizes the binding partner even when the binding partner is present in a mixture of other molecules or organisms. The binding may be mediated by covalent or non-covalent interactions or a combination of both. In yet other words, the term “specifically binding” means immunospecifically binding to an antigenic determinant or epitope and not immunospecifically binding to other antigenic determinants or epitopes.

A binding molecule that immunospecifically binds to an antigen may bind to other peptides or polypeptides with lower affinity as determined by, e.g., radioimmunoassays (RIA), enzyme-linked immunosorbent assays (ELISA), BIACORE, or other assays known in the art. Binding molecules or fragments thereof that immunospecifically bind to an antigen may be cross-reactive with related antigens, carrying the same epitope.

Preferably, binding molecules or fragments thereof that immunospecifically bind to an antigen do not cross-react with other antigens.

A “substitution”, as used herein, denotes the replacement of one or more amino acids or nucleotides by different amino acids or nucleotides, respectively.

The term “therapeutically effective amount” refers to an amount of the binding molecule as defined herein that is effective for preventing, ameliorating and/or treating a condition resulting from infection with an influenza B virus. Ameloriation as used in herein may refer to the reduction of visible or perceptible disease symptoms, viremia, or any other measurable manifestation of influenza infection.

The term “treatment” refers to therapeutic treatment as well as prophylactic or preventative measures to cure or halt or at least retard disease progress. Those in need of treatment include those already inflicted with a condition resulting from infection with influenza virus as well as those in which infection with influenza virus is to be prevented.

Subjects partially or totally recovered from infection with influenza virus might also be in need of treatment. Prevention encompasses inhibiting or reducing the spread of influenza virus or inhibiting or reducing the onset, development or progression of one or more of the symptoms associated with infection with influenza virus.

The term “vector” denotes a nucleic acid molecule into which a second nucleic acid molecule can be inserted for introduction into a host where it will be replicated, and in some cases expressed. In other words, a vector is capable of transporting a nucleic acid molecule to which it has been linked. Cloning as well as expression vectors are contemplated by the term “vector”, as used herein. Vectors include, but are not limited to, plasmids, cosmids, bacterial artificial chromosomes (BAC) and yeast artificial chromosomes (YAC) and vectors derived from bacteriophages or plant or animal (including human) viruses. Vectors comprise an origin of replication recognized by the proposed host and in case of expression vectors, promoter and other regulatory regions recognized by the host. A vector containing a second nucleic acid molecule is introduced into a cell by transformation, transfection, or by making use of viral entry mechanisms.

Certain vectors are capable of autonomous replication in a host into which they are introduced (e.g., vectors having a bacterial origin of replication can replicate in bacteria).

Other vectors can be integrated into the genome of a host upon introduction into the host, and thereby are replicated along with the host genome.

DETAILED DESCRIPTION In a first aspect the present disclosure encompasses binding molecules capable of specifically binding to hemagglutinin (HA) of influenza A virus subtypes of phylogenetic group 1 and influenza A virus subtypes of phylogenetic group 2. In an embodiment, the binding molecules are capable of neutralizing influenza A virus subtypes of both phylogenetic group 1 and phylogenetic group 2. The binding molecules described herein thus are unique in that they are capable of cross-neutralizing group 1 influenza A virus strains and group 2 influenza A virus strains. In an embodiment, the binding molecules are capable of neutralizing at least one or more, preferably two or more, preferably three or more, preferably four or more, even more preferably five or more group 1 influenza A virus subtypes selected from the group consisting of the H1, H2, H5, H6, H8, H9 and H11 subtype, and at least one or more, preferably two or more, preferably three or more group 2 influenza A virus subtypes selected from the group consisting of the H3, H4, H7, and H10 subtype. In an embodiment, the binding molecules are capable of specifically binding to hemagglutinin (HA) of influenza B virus subtypes. In another embodiment, the binding molecules are capable of neutralizing influenza B viruses. In an embodiment, the binding molecules are capable of in vivo neutralizing influenza A and/or B viruses. The influenza A and B virus strains may be both human and non-human influenza virus strains (i.e. obtained from non-human animals, e.g. birds).

Preferably, the binding molecules are human binding molecules. In a preferred embodiment, the binding molecules are human antibodies, or antigen-binding fragments thereof.

In an embodiment, the binding molecules are derived from the VH1-69 germ line gene. Thus, the binding molecules all use the same VH1-69 germ line encoded framework.

In an embodiment, the binding interaction of the binding molecules, preferably the antibody, and HA is mediated exclusively by heavy chain variable sequences.

In an embodiment, the binding molecules comprise a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 133 or SEQ ID NO: 139, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 134, SEQ ID NO: 140 or SEQ ID NO: 151, and a heavy chain CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 135, SEQ ID NO: 141, SEQ ID NO: 145, SEQ ID NO: 152, SEQ ID NO: 161, and SEQ ID NO: 162. The CDR regions of binding molecules described herein are shown in Table 7. CDR regions are according to Kabat et al. (1991) as described in Sequences of Proteins of Immunological Interest.

Influenza viruses infect cells by binding to sialic acid residues on the cell surface of target cells, and following transfer into endosomes, by fusing their membranes with the endosomal membranes and releasing the genome-transcriptase complex into the cell.

Both receptor binding and membrane fusion process are mediated by the HA glycoprotein. The HA of influenza virus A comprises two structurally distinct regions, i.e. a globular head region, which contains a receptor binding site which is responsible for virus attachment to the target cell, and is involved in the haemagglutination activity of HA, and a stem region, containing a fusion peptide which is necessary for membrane fusion between the viral envelope and the endosomal membrane of the cell. The HA protein is a trimer in which each monomer consists of two disulphide - linked glycopolypeptides, HA1 and HA2, that are produced during infection by proteolytic cleavage of a precursor (HA0). Cleavage is necessary for virus infectivity since it is required to prime the HA for membrane fusion, to allow conformational change.

Activation of the primed molecule occurs at low pH in endosomes, between pH5 and pH6, and requires extensive changes in HA structure. Each of the stages in the priming and activation of HA for its participation in the membrane fusion process, presents a different target for inhibition, e.g. by monoclonal antibodies. In an embodiment, the binding molecules are capable of blocking the pH-induced conformational changes in HA associated with membrane fusion.

The binding molecules described herein may be capable of specifically binding to the HA0, HA1 and/or HA2 subunit of the HA protein. They may be capable of specifically binding to linear or structural and/or conformational epitopes on the HA0, HA1 and/or HA2 subunit of the HA protein. The HA molecule may be purified from viruses or recombinantly produced and optionally isolated before use. Alternatively, HA may be expressed on the surface of cells. In an embodiment, the binding molecules described herein are capable of specifically binding to an epitope in the stem region of HA. In an embodiment, the binding molecules bind to an epitope that is accessible in the pre-fusion conformation of HA.

The binding molecules described herein may be capable of specifically binding to influenza viruses that are viable, living and/or infective or that are in inactivated/attenuated form. Methods for inactivating/attenuating virus, e.g. influenza viruses are well known in the art and include, but are not limited to, treatment with formalin, β-propiolactone (BPL), merthiolate, and/or ultraviolet light.

The binding molecules described herein may also be capable of specifically binding to one or more fragments of the influenza viruses, such as inter alia a preparation of one or more proteins and/or (poly)peptides, derived from subtypes of influenza A and/or B viruses or one or more recombinantly produced proteins and/or polypeptides of influenza A and/or B viruses. The nucleotide and/or amino acid sequence of proteins of various influenza A and B strains can be found in the GenBank-database, NCBI Influenza Virus Sequence Database, Influenza Sequence Database (ISD), EMBL-database and/or other databases. It is well within the reach of the skilled person to find such sequences in the respective databases.

In another embodiment the binding molecules described herein are capable of specifically binding to a fragment of the above-mentioned proteins and/or polypeptides, wherein the fragment at least comprises an epitope recognized by the binding molecules described herein. An "epitope" as used herein is a moiety that is capable of binding to a binding molecule described herein with sufficiently high affinity to form a detectable antigen-binding molecule complex.

The binding molecules described herein may or may not be capable of specifically binding to the extracellular part of HA (also called herein soluble HA (sHA)).

The binding molecules described herein can be intact immunoglobulin molecules such as polyclonal or monoclonal antibodies or the binding molecules can be antigen- binding fragments thereof, including, but not limited to, heavy and light chain variable regions, Fab, F(ab'), F(ab') , Fv, dAb, Fd, complementarity determining region (CDR) fragments, single-chain antibodies (scFv), bivalent single-chain antibodies, single-chain phage antibodies, diabodies, triabodies, tetrabodies, and (poly)peptides that contain at least a fragment of an immunoglobulin that is sufficient to confer specific antigen binding to influenza virus strains or a fragment thereof. In a preferred embodiment the binding molecules described herein are human monoclonal antibodies, and/or antigen-binding fragments thereof. The binding molecules may also be Nanobodies, alphabodies, affibodies, FN3-domain scaffolds and other scaffolds based on domains in (human) repeat proteins like Adnectins, Anticalins, Darpins, etc, or other scaffolds comprising epitope binding sequences.

The binding molecules described herein can be used in non-isolated or isolated form. Furthermore, the binding molecules described herein can be used alone or in a mixture comprising at least one binding molecule (or variant or fragment thereof) described herein, and/or with other binding molecules that bind to influenza and have influenza virus inhibiting effect. In other words, the binding molecules can be used in combination, e.g., as a pharmaceutical composition comprising two or more binding molecules of the invention, variants or fragments thereof. For example, binding molecules having different, but complementary activities can be combined in a single therapy to achieve a desired prophylactic, therapeutic or diagnostic effect, but alternatively, binding molecules having identical activities can also be combined in a single therapy to achieve a desired prophylactic, therapeutic or diagnostic effect.

Optionally, the mixture further comprises at least one other therapeutic agent. Preferably, the therapeutic agent such as, e.g., M2 inhibitors (e.g., amantidine, rimantadine) and/or neuraminidase inhibitors (e.g., zanamivir, oseltamivir) is useful in the prophylaxis and/or treatment of an influenza virus infection Typically, binding molecules described herein can bind to their binding partners, i.e. an influenza A virus of group 1 (such as H1N1) and an influenza A virus of group 2 (such as H3N2), and/or an influenza B virus, and/or fragments thereof, with an affinity -4 -5 -6 -7 constant (K -value) that is lower than 0.2x10 M, 1.0x10 M, 1.0x10 M, 1.0x10 M, -8 -9 preferably lower than 1.0x10 M, more preferably lower than 1.0x10 M, more -10 -11 preferably lower than 1.0x10 M, even more preferably lower than 1.0x10 M, and in particular lower than 1.0x10 M. The affinity constants can vary for antibody isotypes.

For example, affinity binding for an IgM isotype refers to a binding affinity of at least about 1.0x10 M. Affinity constants can for instance be measured using surface plasmon resonance, for example using the BIACORE system (Pharmacia Biosensor AB, Uppsala, Sweden).

The binding molecules described herein exhibit neutralizing activity. Neutralizing activity can for instance be measured as described herein. Alternative assays measuring neutralizing activity are described in for instance WHO Manual on Animal Influenza Diagnosis and Surveillance, Geneva: World Health Organisation, 2005, version 2002.5.

Typically, the binding molecules described herein have a neutralizing activity of 50µg/ml or less, preferably 20µg/ml or less, more preferably a neutralizing activity of 10 µg/ml or less, even more preferably 5 µg/ml or less, as determined in an in vitro virus neutralization assay (VNA) as described in Example 6. The binding molecules described herein bind to influenza virus or a fragment thereof in soluble form such as for instance in a sample or in suspension or may bind to influenza viruses or fragments thereof bound or attached to a carrier or substrate, e.g., microtiter plates, membranes and beads, etc.

Carriers or substrates may be made of glass, plastic (e.g., polystyrene), polysaccharides, nylon, nitrocellulose, or Teflon, etc. The surface of such supports may be solid or porous and of any convenient shape. Furthermore, the binding molecules may bind to influenza virus in purified/isolated or non-purified/non-isolated form.

As discussed above, the present disclosure relates to isolated human binding molecules that are able to recognize and bind to an epitope in the influenza haemagglutinin protein (HA) wherein said binding molecules have neutralizing activity against influenza A viruses of phylogenetic group 1 and influenza A viruses of phylogenetic group 2. According to the disclosure, it thus has been shown that the binding molecules of the present invention cross-neutralize influenza virus subtypes belonging to both phylogenetic groups. The skilled person, based on what has been disclosed herein, can determine whether an antibody indeed cross-reacts with HA proteins from different subtypes and can also determine whether they are able to neutralize influenza viruses of different subtypes in vitro and/or in vivo.

In an embodiment the binding molecule according to the present disclosure is selected from the group consisting of: a) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:133, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:135, b) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:140, and a heavy chain CDR3 region of SEQ ID NO:141, c) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, d) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:151, and a heavy chain CDR3 region of SEQ ID NO:152, e) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, f) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:151, and a heavy chain CDR3 region of SEQ ID NO:161, g) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:151, and a heavy chain CDR3 region of SEQ ID NO:162, and h) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:141.

In a preferred embodiment, the binding molecule comprises a heavy chain CDR1 region comprising the amino acid sequence of SEQ ID NO:139, a heavy chain CDR2 region comprising an amino acid sequence of SEQ ID NO:134, and a heavy chain CDR3 region comprising the amino acid sequence of SEQ ID NO:145 or SEQ ID NO: 152.

In another embodiment, the human binding molecules described herein are selected from the group consisting of: a) a binding molecule having a heavy chain CDR1 region of SEQ ID NO:133, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:135, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:136, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:137, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:138, b) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:140, and a heavy chain CDR3 region of SEQ ID NO:141, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:142, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:143, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:144, c) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:146, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:174, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:147, d) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:148, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:149, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:150, e) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:151, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:153, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:154, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:155, f) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:148, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:149, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:150, g) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:156, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:157, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:158, h) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:148, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:159, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:160, i) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:151, and a heavy chain CDR3 region of SEQ ID NO:161, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:142, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:143, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:144, j) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:151, and a heavy chain CDR3 region of SEQ ID NO:162, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:163, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:164, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:165, k) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:166, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:167, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:168, l) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:169, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:149, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:150, m) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:141, a light chain CDR1 region having the amino acid sequence of SEQ ID NO: 163, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:169, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:170, n) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:171, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:164, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:172, o) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:142, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:143, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:173, and p) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:142, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:143, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:144.

In another embodiment, the human binding molecules described herein are selected from the group consisting of: a) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:146, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:174, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:147, b) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:171, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:164, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:172, c) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:145, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:142, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:143, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:173, and d) a binding molecule comprising a heavy chain CDR1 region of SEQ ID NO:139, a heavy chain CDR2 region of SEQ ID NO:134, and a heavy chain CDR3 region of SEQ ID NO:152, a light chain CDR1 region having the amino acid sequence of SEQ ID NO:142, a light chain CDR2 region having the amino acid sequence of SEQ ID NO:143, and a light chain CDR3 region having the amino acid sequence of SEQ ID NO:144.

In another embodiment, the binding molecule described herein is selected from the group consisting of a) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 2, b) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 6, c) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 10, d) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 14, e) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 18, f) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 22, g) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 26, h) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 30, i) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 34, j) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 38, k) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 42, l) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 46, m) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 50, n) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 54, o) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 58, p) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 62.

In an embodiment, the binding molecule according to the invention is selected from the group consisting of a binding molecule comprising a heavy chain variable region of SEQ ID NO: 10 a binding molecule comprising a heavy chain variable region of SEQ ID NO: 54, a binding molecule comprising a heavy chain variable region of SEQ ID NO: 58, and a binding molecule comprising a heavy chain variable region of SEQ ID NO: 62.

In a further embodiment, the binding molecules described herein comprise a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:4, SEQ ID NO:8, SEQ ID NO:12, SEQ ID NO:16, SEQ ID NO:20, SEQ ID NO:24, SEQ ID NO:28, SEQ ID NO:32, SEQ ID NO:36, SEQ ID NO:40, SEQ ID NO:44, SEQ ID NO:48, SEQ ID NO:52, SEQ ID NO:56, SEQ ID NO:60, and SEQ ID NO:64.

In yet another embodiment, the binding molecule is selected from the group consisting of a) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 2 and a light chain variable region of SEQ ID NO: 4, b) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 6 and a light chain variable region of SEQ ID NO: 8, c) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 10 and a light chain variable region of SEQ ID NO: 12, d) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 14 and a light chain variable region of SEQ ID NO: 16, e) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 18 and a light chain variable region of SEQ ID NO: 20, f) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 22 and a light chain variable region of SEQ ID NO: 24, g) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 26 and a light chain variable region of SEQ ID NO: 28, h) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 30 and a light chain variable region of SEQ ID NO: 32, i) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 34 and a light chain variable region of SEQ ID NO: 36, j) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 38 and a light chain variable region of SEQ ID NO: 40, k) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 42 and a light chain variable region of SEQ ID NO: 44, l) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 46 and a light chain variable region of SEQ ID NO: 48, m) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 50 and a light chain variable region of SEQ ID NO: 52, n) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 54 and a light chain variable region of SEQ ID NO: 56, o) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 58 and a light chain variable region of SEQ ID NO: 60, and p) a binding molecule comprising a heavy chain variable region of SEQ ID NO: 62 and a light chain variable region of SEQ ID NO: 64.

In an embodiment, the human binding molecules described herein are selected from the group consisting of: a binding molecule comprising a heavy chain variable region of SEQ ID NO: 10 and a light chain variable region of SEQ ID NO: 12, a binding molecule comprising a heavy chain variable region of SEQ ID NO: 54 and a light chain variable region of SEQ ID NO: 56, a binding molecule comprising a heavy chain variable region of SEQ ID NO: 58 and a light chain variable region of SEQ ID NO: 60, and a binding molecule comprising a heavy chain variable region of SEQ ID NO: 62 and a light chain variable region of SEQ ID NO: 64.

In a preferred embodiment, the binding molecules described herein are for a use as a medicament, and preferably for use in the diagnostic, therapeutic and/or prophylactic treatment of influenza infection caused by influenza A and/or B viruses. Preferably, the influenza virus that causes the influenza infection and that can be treated using the binding molecules of the present invention is an influenza A virus of phylogenetic group 1 and/or 2, and/or a influenza B virus. Also described is a pharmaceutical composition comprising at least one binding molecule according to the invention, and a pharmaceutically acceptable excipient.

Also described is the use of a binding molecule according to the invention in the preparation of a medicament for the diagnosis, prophylaxis, and/or treatment of an influenza virus infection. Such infections can occur in small populations, but can also spread around the world in seasonal epidemics or, worse, in global pandemics where millions of individuals are at risk. Also described are binding molecules that can neutralize the infection of influenza strains that cause such seasonal epidemics, as well as potential pandemics. Importantly, protection and treatment can be envisioned now with the binding molecules described herein in relation to various influenza subtypes as it has been disclosed that the binding molecules of the present invention are capable of cross- neutralizing various influenza subtypes of both phylogenetic group 1, encompassing H1, H2, H5, H6, H8, H9 and H11 subtypes and phylogenetic group 2, encompassing subtypes H3, H4, H7 and H10 subtypes, as well as influenza B subtypes.

Also described are functional variants of the binding molecules as defined herein.

Molecules are considered to be functional variants of a binding molecule according to the invention, if the variants are capable of competing for specifically binding to an influenza virus or a fragment thereof with the “parental” or “reference” binding molecules. In other words, molecules are considered to be functional variants of a binding molecule described herein when the functional variants are still capable of binding to the same or overlapping epitope of the influenza virus or a fragment thereof. For the sake of this application “parental” and “reference” will be used as synonyms meaning that the information of the reference or parental molecule, or the physical molecule itself may form the basis for the variation. Functional variants include, but are not limited to, derivatives that are substantially similar in primary structural sequence, including those that have modifications in the Fc receptor or other regions involved with effector functions, and/or which contain e.g. in vitro or in vivo modifications, chemical and/or biochemical, that are not found in the parental binding molecule. Such modifications include inter alia acetylation, acylation, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, cross-linking, disulfide bond formation, glycosylation, hydroxylation, methylation, oxidation, pegylation, proteolytic processing, phosphorylation, and the like.

Alternatively, functional variants can be binding molecules as defined herein comprising an amino acid sequence containing substitutions, insertions, deletions or combinations thereof of one or more amino acids compared to the amino acid sequences of the parental binding molecules. Furthermore, functional variants can comprise truncations of the amino acid sequence at either or both the amino or carboxyl termini. Functional variants described herein may have the same or different, either higher or lower, binding affinities compared to the parental binding molecule but are still capable of binding to the influenza virus or a fragment thereof. For instance, functional variants described herein may have increased or decreased binding affinities for an influenza virus or a fragment thereof compared to the parental binding molecules. Preferably, the amino acid sequences of the variable regions, including, but not limited to, framework regions, hypervariable regions, in particular the CDR3 regions, are modified. Generally, the light chain and the heavy chain variable regions comprise three hypervariable regions, comprising three CDRs, and more conserved regions, the so-called framework regions (FRs). The hypervariable regions comprise amino acid residues from CDRs and amino acid residues from hypervariable loops. Functional variants intended to fall within the scope of the present disclosure have at least about 50% to about 99%, preferably at least about 60% to about 99%, more preferably at least about 70% to about 99%, even more preferably at least about 80% to about 99%, most preferably at least about 90% to about 99%, in particular at least about 95% to about 99%, and in particular at least about 97% to about 99% amino acid sequence identity and/or homology with the parental binding molecules as defined herein. Computer algorithms such as inter alia Gap or Bestfit known to a person skilled in the art can be used to optimally align amino acid sequences to be compared and to define similar or identical amino acid residues. Functional variants can be obtained by altering the parental binding molecules or parts thereof by general molecular biology methods known in the art including, but not limited to, error-prone PCR, oligonucleotide-directed mutagenesis, site-directed mutagenesis and heavy and/or light chain shuffling. In an embodiment the functional variants described herein have neutralizing activity against influenza A viruses of group 1 and group 2, and/or influenza B viruses. The neutralizing activity may either be identical, or be higher or lower compared to the parental binding molecules. Henceforth, when the term (human) binding molecule is used, this also encompasses functional variants of the (human) binding molecule. Assays for verifying if a variant binding molecule has neutralizing activity are well known in the art (see WHO Manual on Animal Influenza Diagnosis and Surveillance, Geneva: World Health Organisation, 2005 version 2002.5).

Also described are immunoconjugates, i.e. molecules comprising at least one binding molecule as defined herein and further comprising at least one tag, such as inter alia a detectable moiety/agent. Also contemplated herein are mixtures of immunoconjugates described herein or mixtures of at least one immunoconjugates described herein and another molecule, such as a therapeutic agent or another binding molecule or immunoconjugate. In a further embodiment, the immunoconjugates described herein may comprise more than one tag. These tags can be the same or distinct from each other and can be joined/conjugated non-covalently to the binding molecules.

The tag(s) can also be joined/conjugated directly to the human binding molecules through covalent bonding. Alternatively, the tag(s) can be joined/conjugated to the binding molecules by means of one or more linking compounds. Techniques for conjugating tags to binding molecules are well known to the skilled artisan.

The tags of the immunoconjugates described herein may be therapeutic agents, but they can also be detectable moieties/agents. Tags suitable in therapy and/or prevention may be toxins or functional parts thereof, antibiotics, enzymes, other binding molecules that enhance phagocytosis or immune stimulation. Immunoconjugates comprising a detectable agent can be used diagnostically to, for example, assess if a subject has been infected with an influenza virus or to monitor the development or progression of an influenza virus infection as part of a clinical testing procedure to, e.g., determine the efficacy of a given treatment regimen. However, they may also be used for other detection and/or analytical and/or diagnostic purposes. Detectable moieties/agents include, but are not limited to, enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron emitting metals, and non-radioactive paramagnetic metal ions. The tags used to label the binding molecules for detection and/or analytical and/or diagnostic purposes depend on the specific detection/analysis/diagnosis techniques and/or methods used such as inter alia immunohistochemical staining of (tissue) samples, flow cytometric detection, scanning laser cytometric detection, fluorescent immunoassays, enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), bioassays (e.g., phagocytosis assays), Western blotting applications, etc. Suitable labels for the detection/analysis/diagnosis techniques and/or methods known in the art are well within the reach of the skilled artisan.

Furthermore, the human binding molecules or immunoconjugates described herein can also be attached to solid supports, which are particularly useful for in vitro immunoassays or purification of influenza viruses or fragments thereof. Such solid supports might be porous or nonporous, planar or non-planar. The binding molecules described herein can be fused to marker sequences, such as a peptide to facilitate purification. Examples include, but are not limited to, the hexa-histidine tag, the hemagglutinin (HA) tag, the myc tag or the flag tag. Alternatively, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate. In another aspect the binding molecules described herein may be conjugated/attached to one or more antigens. Preferably, these antigens are antigens which are recognized by the immune system of a subject to which the binding molecule-antigen conjugate is administered. The antigens may be identical, but may also differ from each other. Conjugation methods for attaching the antigens and binding molecules are well known in the art and include, but are not limited to, the use of cross-linking agents. The binding molecules described herein will bind to influenza virus HA and the antigens attached to the binding molecules will initiate a powerful T-cell attack on the conjugate, which will eventually lead to the destruction of the influenza virus.

Next to producing immunoconjugates chemically by conjugating, directly or indirectly, via for instance a linker, the immunoconjugates can be produced as fusion proteins comprising the binding molecules described herein and a suitable tag. Fusion proteins can be produced by methods known in the art such as, e.g., recombinantly by constructing nucleic acid molecules comprising nucleotide sequences encoding the binding molecules in frame with nucleotide sequences encoding the suitable tag(s) and then expressing the nucleic acid molecules.

Also described is a nucleic acid molecule encoding at least a binding molecule, functional variant or immunoconjugate described herein. Such nucleic acid molecules can be used as intermediates for cloning purposes, e.g. in the process of affinity maturation as described above. In a preferred embodiment, the nucleic acid molecules are isolated or purified.

The skilled man will appreciate that functional variants of these nucleic acid molecules are also intended to be a part of the present disclosure. Functional variants are nucleic acid sequences that can be directly translated, using the standard genetic code, to provide an amino acid sequence identical to that translated from the parental nucleic acid molecules.

Preferably, the nucleic acid molecules encode binding molecules comprising the CDR regions as described above. In a further embodiment the nucleic acid molecules encode binding molecules comprising two, three, four, five or even all six CDR regions of the binding molecules described herein.

In another embodiment, the nucleic acid molecules encode binding molecules comprising a heavy chain comprising the variable heavy chain sequences as described above. In another embodiment the nucleic acid molecules encode binding molecules comprising a light chain comprising the variable light chain sequences as described above. The nucleotide sequences and the amino acid sequences of the heavy and light chain variable regions of the binding molecules described herein are given below.

Also described are vectors, i.e. nucleic acid constructs, comprising one or more nucleic acid molecules described herein. Vectors can be derived from plasmids such as inter alia F, R1, RP1, Col, pBR322, TOL, Ti, etc; cosmids; phages such as lambda, lambdoid, M13, Mu, P1, P22, Q β, T-even, T-odd, T2, T4, T7, etc; plant viruses. Vectors can be used for cloning and/or for expression of the binding molecules of the invention and might even be used for gene therapy purposes. Vectors comprising one or more nucleic acid molecules described herein operably linked to one or more expression- regulating nucleic acid molecules are also covered by the present disclosure. The choice of the vector is dependent on the recombinant procedures followed and the host used.

Introduction of vectors in host cells can be effected by inter alia calcium phosphate transfection, virus infection, DEAE-dextran mediated transfection, lipofectamin transfection or electroporation. Vectors may be autonomously replicating or may replicate together with the chromosome into which they have been integrated. Preferably, the vectors contain one or more selection markers. The choice of the markers may depend on the host cells of choice, although this is not critical to the invention as is well known to persons skilled in the art. They include, but are not limited to, kanamycin, neomycin, puromycin, hygromycin, zeocin, thymidine kinase gene from Herpes simplex virus (HSV-TK), dihydrofolate reductase gene from mouse (dhfr). Vectors comprising one or more nucleic acid molecules encoding the human binding molecules as described above operably linked to one or more nucleic acid molecules encoding proteins or peptides that can be used to isolate the human binding molecules are also covered by the disclosure.

These proteins or peptides include, but are not limited to, glutathione-S-transferase, maltose binding protein, metal-binding polyhistidine, green fluorescent protein, luciferase and beta-galactosidase.

Hosts containing one or more copies of the vectors mentioned above are an additional subject of the present disclosure. Preferably, the hosts are host cells. Host cells include, but are not limited to, cells of mammalian, plant, insect, fungal or bacterial origin. Bacterial cells include, but are not limited to, cells from Gram-positive bacteria or Gram-negative bacteria such as several species of the genera Escherichia, such as E. coli, and Pseudomonas. In the group of fungal cells preferably yeast cells are used. Expression in yeast can be achieved by using yeast strains such as inter alia Pichia pastoris, Saccharomyces cerevisiae and Hansenula polymorpha. Furthermore, insect cells such as cells from Drosophila and Sf9 can be used as host cells. Besides that, the host cells can be plant cells such as inter alia cells from crop plants such as forestry plants, or cells from plants providing food and raw materials such as cereal plants, or medicinal plants, or cells from ornamentals, or cells from flower bulb crops. Transformed (transgenic) plants or plant cells are produced by known methods, for example, Agrobacterium-mediated gene transfer, transformation of leaf discs, protoplast transformation by polyethylene glycol- induced DNA transfer, electroporation, sonication, microinjection or bolistic gene transfer. Additionally, a suitable expression system can be a baculovirus system.

Expression systems using mammalian cells, such as Chinese Hamster Ovary (CHO) cells, COS cells, BHK cells, NSO cells or Bowes melanoma cells are preferred herein.

Mammalian cells provide expressed proteins with posttranslational modifications that are most similar to natural molecules of mammalian origin. Since the present disclosure deals with molecules that may have to be administered to humans, a completely human expression system would be particularly preferred. Therefore, even more preferably, the host cells are human cells. Examples of human cells are inter alia HeLa, 911, AT1080, A549, 293 and HEK293T cells. In preferred embodiments, the human producer cells comprise at least a functional part of a nucleic acid sequence encoding an adenovirus E1 region in expressible format. In even more preferred embodiments, said host cells are derived from a human retina and immortalized with nucleic acids comprising adenoviral E1 sequences, such as 911 cells or the cell line deposited at the European Collection of Cell Cultures (ECACC), CAMR, Salisbury, Wiltshire SP4 OJG, Great Britain on 29 February 1996 under number 96022940 and marketed under the trademark PER.C6 (PER.C6 is a registered trademark of Crucell Holland B.V.). For the purposes of this application “PER.C6 cells” refers to cells deposited under number 96022940 or ancestors, passages up-stream or downstream as well as descendants from ancestors of deposited cells, as well as derivatives of any of the foregoing. Production of recombinant proteins in host cells can be performed according to methods well known in the art. The use of the cells marketed under the trademark PER.C6 as a production platform for proteins of interest has been described in WO 00/63403 the disclosure of which is incorporated herein by reference in its entirety.

In yet another embodiment, binding molecules described herein can also be produced in transgenic, non-human, mammals such as inter alia rabbits, goats or cows, and secreted into for instance the milk thereof.

In yet another alternative embodiment, binding molecules described herein may be generated by transgenic non-human mammals, such as for instance transgenic mice or rabbits that express human immunoglobulin genes. Preferably, the transgenic non-human mammals have a genome comprising a human heavy chain transgene and a human light chain transgene encoding all or a portion of the human binding molecules as described above. The transgenic non-human mammals can be immunized with a purified or enriched preparation of influenza virus or a fragment thereof. Protocols for immunizing non-human mammals are well established in the art. See Using Antibodies: A Laboratory Manual, Edited by: E. Harlow, D. Lane (1998), Cold Spring Harbor Laboratory, Cold Spring Harbor, New York and Current Protocols in Immunology, Edited by: J.E. Coligan, A.M. Kruisbeek, D.H. Margulies, E.M. Shevach, W. Strober (2001), John Wiley & Sons Inc., New York, the disclosures of which are incorporated herein by reference.

Immunization protocols often include multiple immunizations, either with or without adjuvants such as Freund's complete adjuvant and Freund's incomplete adjuvant, but may also include naked DNA immunizations. In another embodiment, the human binding molecules are produced by B-cells, plasma and/or memory cells derived from the transgenic animals. In yet another embodiment, the human binding molecules are produced by hybridomas, which are prepared by fusion of B-cells obtained from the above-described transgenic non-human mammals to immortalized cells. B-cells, plasma cells and hybridomas as obtainable from the above-described transgenic non-human mammals and human binding molecules as obtainable from the above-described transgenic non-human mammals, B-cells, plasma and/or memory cells and hybridomas are also a part of the present disclosure.

Also described are compositions comprising at least a binding molecule, preferably a human monoclonal antibody, described herein, at least a functional variant thereof, at least an immunoconjugate described herein and/or a combination thereof. In addition to that, the compositions may comprise inter alia stabilizing molecules, such as albumin or polyethylene glycol, or salts. Preferably, the salts used are salts that retain the desired biological activity of the binding molecules and do not impart any undesired toxicological effects. If necessary, the human binding molecules described herein may be coated in or on a material to protect them from the action of acids or other natural or non- natural conditions that may inactivate the binding molecules.

Also described are compositions comprising at least a nucleic acid molecule as defined herein. The compositions may comprise aqueous solutions such as aqueous solutions containing salts (e.g., NaCl or salts as described above), detergents (e.g., SDS) and/or other suitable components.

Also described are pharmaceutical compositions comprising at least a binding molecule such as a human monoclonal antibody described herein (or functional fragment or variant thereof), at least an immunoconjugate described herein, at least a composition described herein, or combinations thereof. The pharmaceutical composition described herein further comprises at least one pharmaceutically acceptable excipient.

Pharmaceutically acceptable excipients are well known to the skilled person. The pharmaceutical composition described herein may further comprise at least one other therapeutic agent. Suitable agents are also well known to the skilled artisan.

In a preferred embodiment the pharmaceutical composition described herein comprises at least one additional binding molecule, i.e. the pharmaceutical composition can be a cocktail or mixture of binding molecules. The pharmaceutical composition may comprise at least two binding molecules described herein, or at least one binding molecule described herein and at least one further influenza virus binding and/or neutralizing molecule, such as another antibody directed against the HA protein or against other antigenic structures present on influenza viruses, such as M2. In another embodiment the additional binding molecule may be formulated for simultaneous separate or sequential administration.

In an embodiment the pharmaceutical compositions may comprise two or more binding molecules that have neutralizing activity against influenza A viruses and/or influenza B viruses. In an embodiment, the binding molecules exhibit synergistic neutralizing activity, when used in combination. As used herein, the term "synergistic" means that the combined effect of the binding molecules when used in combination is greater than their additive effects when used individually. The synergistically acting binding molecules may bind to different structures on the same or distinct fragments of influenza virus. A way of calculating synergy is by means of the combination index. The concept of the combination index (CI) has been described by Chou and Talalay (1984).

The compositions may e.g. comprise one binding molecule having neutralizing activity and one non-neutralizing binding molecule. The non-neutralizing and neutralizing binding molecules may also act synergistically in neutralizing influenza virus.

In an embodiment, the pharmaceutical composition may comprise at least one binding molecule described herein and at least one further influenza virus neutralizing binding molecule. The binding molecules in the pharmaceutical composition preferably are capable of reacting with influenza viruses of different subtypes. The binding molecules should be of high affinity and should have a broad specificity. Preferably, both binding molecules are cross-neutralizing molecules in that they each neutralize influenza viruses of different subtypes. In addition, preferably they neutralize as many strains of each of the different influenza virus subtypes as possible.

A pharmaceutical composition described herein can further comprise at least one other therapeutic, prophylactic and/or diagnostic agent. Preferably, the pharmaceutical composition comprises at least one other prophylactic and/or therapeutic agent.

Preferably, said further therapeutic and/or prophylactic agents are agents capable of preventing and/or treating an influenza virus infection and/or a condition resulting from such an infection. Therapeutic and/or prophylactic agents include, but are not limited to, anti-viral agents. Such agents can be binding molecules, small molecules, organic or inorganic compounds, enzymes, polynucleotide sequences, anti-viral peptides, etc. Other agents that are currently used to treat patients infected with influenza viruses are M2 inhibitors (e.g., amantidine, rimantadine) and/or neuraminidase inhibitors (e.g., zanamivir, oseltamivir). These can be used in combination with the binding molecules described herein. “In combination” herein means simultaneously, as separate formulations, or as one single combined formulation, or according to a sequential administration regimen as separate formulations, in any order. Agents capable of preventing and/or treating an infection with influenza virus and/or a condition resulting from such an infection that are in the experimental phase might also be used as other therapeutic and/or prophylactic agents useful herein.

The binding molecules or pharmaceutical compositions described herein can be tested in suitable animal model systems prior to use in humans. Such animal model systems include, but are not limited to, mouse, ferret and monkey.

Typically, pharmaceutical compositions must be sterile and stable under the conditions of manufacture and storage. The binding molecules, immunoconjugates, nucleic acid molecules or compositions described herein can be in powder form for reconstitution in the appropriate pharmaceutically acceptable excipient before or at the time of delivery. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying (lyophilization) that yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Alternatively, the binding molecules, immunoconjugates, nucleic acid molecules or compositions described herein can be in solution and the appropriate pharmaceutically acceptable excipient can be added and/or mixed before or at the time of delivery to provide a unit dosage injectable form. Preferably, the pharmaceutically acceptable excipient used herein is suitable to high drug concentration, can maintain proper fluidity and, if necessary, can delay absorption.

The choice of the optimal route of administration of the pharmaceutical compositions will be influenced by several factors including the physicochemical properties of the active molecules within the compositions, the urgency of the clinical situation and the relationship of the plasma concentrations of the active molecules to the desired therapeutic effect. For instance, if necessary, the binding molecules described herein can be prepared with carriers that will protect them against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can inter alia be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Furthermore, it may be necessary to coat the binding molecules with, or co-administer the binding molecules with, a material or compound that prevents the inactivation of the human binding molecules. For example, the binding molecules may be administered to a subject in an appropriate carrier, for example, liposomes or a diluent.

The routes of administration can be divided into two main categories, oral and parenteral administration. The preferred administration route is intravenous or by inhalation.

Oral dosage forms can be formulated inter alia as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard capsules, soft gelatin capsules, syrups or elixirs, pills, dragees, liquids, gels, or slurries. These formulations can contain pharmaceutically excipients including, but not limited to, inert diluents, granulating and disintegrating agents, binding agents, lubricating agents, preservatives, colouring, flavouring or sweetening agents, vegetable or mineral oils, wetting agents, and thickening agents.

The pharmaceutical compositions described herein can also be formulated for parenteral administration. Formulations for parenteral administration can be inter alia in the form of aqueous or non-aqueous isotonic sterile non-toxic injection or infusion solutions or suspensions. The solutions or suspensions may comprise agents that are non- toxic to recipients at the dosages and concentrations employed such as 1,3-butanediol, Ringer’s solution, Hank’s solution, isotonic sodium chloride solution, oils, fatty acids, local anaesthetic agents, preservatives, buffers, viscosity or solubility increasing agents, water-soluble antioxidants, oil-soluble antioxidants and metal chelating agents.

In a further aspect, the binding molecules such as human monoclonal antibodies (functional fragments and variants thereof), immunoconjugates, compositions, or pharmaceutical compositions described herein can be used as a medicament. So, a method of diagnosis, treatment and/or prevention of an influenza virus infection using the binding molecules, immunoconjugates, compositions, or pharmaceutical compositions described herein is another part of the present disclosure. The above-mentioned molecules can inter alia be used in the diagnosis, prophylaxis, treatment, or combination thereof, of an influenza virus infection caused influenza viruses comprising HA of the H1, H2, H3, H4, H5, H6, H7, H8, H9, H10 and/or H11 subtype. In an embodiment, the above-mentioned molecules can also be used in the diagnosis, prophylaxis, treatment or combination thereof of an influenza virus infection caused by an influenza B virus. They are suitable for treatment of yet untreated patients suffering from an influenza virus infection and patients who have been or are treated for an influenza virus infection.

The above-mentioned molecules or compositions may be employed in conjunction with other molecules useful in diagnosis, prophylaxis and/or treatment. They can be used in vitro, ex vivo or in vivo. For instance, the binding molecules such as human monoclonal antibodies (or functional variants thereof), immunoconjugates, compositions or pharmaceutical compositions described herein can be co-administered with a vaccine against influenza virus (if available). Alternatively, the vaccine may also be administered before or after administration of the molecules described herein. Instead of a vaccine, anti-viral agents can also be employed in conjunction with the binding molecules of the present invention. Suitable anti-viral agents are mentioned above.

The molecules are typically formulated in the compositions and pharmaceutical compositions described herein in a therapeutically or diagnostically effective amount.

Alternatively, they may be formulated and administered separately. For instance the other molecules such as the anti-viral agents may be applied systemically, while the binding molecules described herein may be applied intravenously.

Treatment may be targeted at patient groups that are susceptible to influenza infection. Such patient groups include, but are not limited to e.g., the elderly (e.g. ≥ 50 years old, ≥ 60 years old, and preferably ≥ 65 years old), the young (e.g. ≤ 5 years old, ≤ 1 year old), hospitalized patients and already infected patients who have been treated with an antiviral compound but have shown an inadequate antiviral response.

Dosage regimens can be adjusted to provide the optimum desired response (e.g., a therapeutic response). A suitable dosage range may for instance be 0.01-100 mg/kg body weight, preferably 0.1-50 mg/kg body weight, preferably 0.01-15 mg/kg body weight. Furthermore, for example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. The molecules and compositions described herein are preferably sterile. Methods to render these molecules and compositions sterile are well known in the art. The other molecules useful in diagnosis, prophylaxis and/or treatment can be administered in a similar dosage regimen as proposed for the binding molecules described herein. If the other molecules are administered separately, they may be administered to a patient prior to (e.g., 2 min, 5 min, 10 min, 15 min, 30 min, 45 min, 60 min, 2 hrs, 4 hrs, 6 hrs, 8 hrs, 10 hrs, 12 hrs, 14 hrs, 16 hrs, 18 hrs, 20 hrs, 22 hrs, 24 hrs, 2 days, 3 days, 4 days, 5 days, 7 days, 2 weeks, 4 weeks or 6 weeks before), concomitantly with, or subsequent to (e.g., 2 min, 5 min, 10 min, 15 min, 30 min, 45 min, 60 min, 2 hrs, 4 hrs, 6 hrs, 8 hrs, 10 hrs, 12 hrs, 14 hrs, 16 hrs, 18 hrs, 20 hrs, 22 hrs, 24 hrs, 2 days, 3 days, 4 days, 5 days, 7 days, 2 weeks, 4 weeks or 6 weeks after) the administration of one or more of the human binding molecules or pharmaceutical compositions described herein. The exact dosing regimen is usually sorted out during clinical trials in human patients.

Human binding molecules and pharmaceutical compositions comprising the human binding molecules are particularly useful, and often preferred, when to be administered to human beings as in vivo therapeutic agents, since recipient immune response to the administered antibody will often be substantially less than that occasioned by administration of a monoclonal murine, chimeric or humanized binding molecule.

Also described is the use of the binding molecules such as neutralizing human monoclonal antibodies (functional fragments and variants thereof), immunoconjugates, nucleic acid molecules, compositions or pharmaceutical compositions described herein in the preparation of a medicament for the diagnosis, prophylaxis, treatment, or combination thereof, of an influenza virus infection, in particular an influenza virus infection caused influenza viruses comprising HA of the H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, and/or H11 subtype and/or influenza B viruses.

Next to that, kits comprising at least a binding molecule such as a neutralizing human monoclonal antibody (functional fragments and variants thereof), at least an immunoconjugate, at least a nucleic acid molecule, at least a composition, at least a pharmaceutical composition, at least a vector, at least a host described herein or a combination thereof are also a part of the present disclosure. Optionally, the above- described components of the kits described herein are packed in suitable containers and labelled for diagnosis, prophylaxis and/or treatment of the indicated conditions. The above-mentioned components may be stored in unit or multi-dose containers as an aqueous, preferably sterile, solution or as a lyophilised, preferably sterile, formulation for reconstitution. The containers may be formed from a variety of materials such as glass or plastic and may have a sterile access port (for example, the container may be an intravenous solution bag or a vial having a stopper pierceable by a hypodermic injection needle). The kit may further comprise more containers comprising a pharmaceutically acceptable buffer. It may further include other materials desirable from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, culture medium for one or more of the suitable hosts and, possibly, even at least one other therapeutic, prophylactic or diagnostic agent. Associated with the kits can be instructions customarily included in commercial packages of therapeutic, prophylactic or diagnostic products, that contain information about for example the indications, usage, dosage, manufacture, administration, contra-indications and/or warnings concerning the use of such therapeutic, prophylactic or diagnostic products.

The binding molecules described herein can also be advantageously used as a diagnostic agent in an in vitro method for the detection of influenza virus. The disclosure thus further pertains to a method of detecting influenza virus phylogenetic group 1 or group 2, or influenza B subtype influenza virus in a sample, wherein the method comprises the steps of (a) contacting a sample with a diagnostically effective amount of a binding molecule (functional fragments and variants thereof) or an immunoconjugate described herein, and (b) determining whether the binding molecule or immunoconjugate specifically binds to a molecule of the sample. The sample may be a biological sample including, but not limited to blood, serum, stool, sputum, nasophargyal aspirates, bronchial lavages, urine, tissue or other biological material from (potentially) infected subjects, or a non-biological sample such as water, drink, etc. The (potentially) infected subjects may be human subjects, but also animals that are suspected as carriers of influenza virus might be tested for the presence of the virus using the human binding molecules or immunoconjugates described herein. The sample may first be manipulated to make it more suitable for the method of detection. Manipulation means inter alia treating the sample suspected to contain and/or containing the virus in such a way that the virus will disintegrate into antigenic components such as proteins, (poly)peptides or other antigenic fragments. Preferably, the human binding molecules or immunoconjugates described herein are contacted with the sample under conditions which allow the formation of an immunological complex between the human binding molecules and the virus or antigenic components thereof that may be present in the sample. The formation of an immunological complex, if any, indicating the presence of the virus in the sample, is then detected and measured by suitable means. Such methods include, inter alia, homogeneous and heterogeneous binding immunoassays, such as radio-immunoassays (RIA), ELISA, immunofluorescence, immunohistochemistry, FACS, BIACORE and Western blot analyses.

Preferred assay techniques, especially for large-scale clinical screening of patient sera and blood and blood-derived products are ELISA and Western blot techniques.

ELISA tests are particularly preferred. For use as reagents in these assays, the binding molecules or immunoconjugates described herein are conveniently bonded to the inside surface of microtiter wells. The binding molecules or immunoconjugates described herein may be directly bonded to the microtiter well. However, maximum binding of the binding molecules or immunoconjugates described herein to the wells might be accomplished by pre-treating the wells with polylysine prior to the addition of the binding molecules or immunoconjugates described herein. Furthermore, the binding molecules or immunoconjugates described herein may be covalently attached by known means to the wells. Generally, the binding molecules or immunoconjugates are used in a concentration between 0.01 to 100 μg/ml for coating, although higher as well as lower amounts may also be used. Samples are then added to the wells coated with the binding molecules or immunoconjugates described herein.

Furthermore, binding molecules described herein can be used to identify specific binding structures of influenza virus. The binding structures can be epitopes on proteins and/or polypeptides. They can be linear, but also structural and/or conformational. In one embodiment, the binding structures can be analysed by means of PEPSCAN analysis (see inter alia WO 84/03564, WO 93/09872, Slootstra et al., 1996). Alternatively, a random peptide library comprising peptides from a protein of influenza virus can be screened for peptides capable of binding to the binding molecules described herein.

The invention is further illustrated in the following examples and figures. The examples are not intended to limit the scope of the invention in any way.

EXAMPLES Example 1 Construction of scFv phage display libraries using RNA extracted from peripheral blood mononuclear cells Peripheral blood was collected from normal healthy donors by venapuncture in EDTA anti-coagulation sample tubes. scFv phage display libraries were obtained as described in , which is incorporated by reference herein. RNA was isolated from peripheral blood mononuclear cells and cDNA prepared. A two round PCR amplification approach was applied using the primer sets shown in Tables 1 and 2 to isolate the immunoglobulin VH and VL regions from the respective donor repertoire.

First round amplification on the respective cDNA using the primer sets mentioned in Table 1 yielded 7, 6 and 9 products of about 650 base pairs for respectively VH, Vkappa and Vlambda regions. For IgM VH region amplification the OCM constant primer was used in combination with OH1 to OH7. The thermal cycling program for first round amplifications was: 2 min 96°C (denaturation step), 30 cycles of 30 sec 96°C/ 30 sec 60°C/ 60 sec 72°C, 10 min 72°C final elongation and 6°C refrigeration. The products were loaded on and isolated from a 1% agarose gel using gel-extraction columns (Macherey Nagel) and eluted in 50 μl 5 mM Tris-HCl pH 8.0. Ten percent of first round products (3 to 5 μl) was subjected to second round amplification using the primers mentioned in Table 2. These primers were extended with restriction sites enabling the directional cloning of the respective VL and VH regions into phage display vector PDV- C06. The PCR program for second round amplifications was as follows: 2 min 96°C (denaturation step), 30 cycles of 30 sec 96°C/ 30 sec 60°C/ 60 sec 72°C, 10 min 72°C final elongation and 6°C refrigeration. The second round products (~350 base pairs) were first pooled according to natural occurrence of J segments found in immunoglobulin gene products, resulting in 7, 6 and 9 pools for respectively the VH, Vkappa and Vlambda variable regions (see Tables 3 and 4). To obtain a normalized distribution of immunoglobulin sequences in the immune library the 6 Vkappa and 9 Vlambda light chain pools were mixed according to the percentages mentioned in Table 3. This single final VL pool (3 μg) was digested overnight with SalI and NotI restriction enzymes, loaded on and isolated from a 1% agarose gel (~350 base pairs) using Macherey Nagel gel-extraction columns and ligated in SalI-NotI cut PDV-C06 vector (~5000 base pairs) as follows: 10 μl PDV-C06 vector (50 ng/ μl), 7 μl VL insert (10 ng/ μl), 5 μl 10X ligation buffer (NEB), 2.5 T4 DNA Ligase (400 U/ μl) (NEB), 25.5 μl ultrapure water (vector to insert ratio was 1:2). Ligation was performed overnight in a water bath of 16°C. Next, the volume was doubled with water, extracted with an equal volume of phenol-chloroform- isoamylalcohol (75:24:1) (Invitrogen) followed by chloroform (Merck) extraction and precipitated with 1 μl Pellet Paint (Novogen), 10 μl sodium acetate (3 M pH 5.0) and 100 μl isopropanol for 2 hrs at -20°C. The obtained sample was subsequently centrifuged at .000xg for 30 min at 4°C. The obtained precipitate was washed with 70% ethanol and centrifuged for 10 min at 20.000xg at room temperature. Ethanol was removed by vacuum aspiration and the pellet was air dried for several min and then dissolved in 50 μl buffer containing 10 mM Tris-HCl, pH 8.0. 2 μl ligation mixture was used for the transformation of 40 μl TG-1 electro-competent cells (Agilent) in a chilled 0.1 cm electroporation cuvette (Biorad) using a Genepulser II apparatus (Biorad) set at 1.7 kV, 200 Ohm, 25 μF (time constant ~4,5 msec). Directly after pulse, the bacteria were flushed from the cuvette with 1000 μl SOC medium (Invitrogen) containing 5% (w/v) glucose (Sigma) at 37°C and transferred to a 15 ml round bottom culture tube. Another 500 μl SOC/glucose was used to flush residual bacteria from the cuvette and was added to the culture tube. Bacteria were recovered by culturing for exactly one hr at 37°C in a shaker incubator at 220 rpm. The transformed bacteria were plated over large 240 mm square petridishes (NUNC) containing 150 ml 2TY agar (16 g/l bacto-tryptone, 10 g/l bacto- yeast extract, 5 g/l NaCl, 15 g/l agar, pH 7.0) supplemented with 50 μg/ml ampicillin and % (w/v) glucose (Sigma). A 1 to 1000 dilution was plated for counting purposes on 15 cm petridishes containing the same medium. This transformation procedure was repeated sequentially ten times and the complete each transformation was plated on a separate square petridish and grown overnight in a 37°C culture stove. Typically, around 1x10 cfu (1x10 per petridish) were obtained using the above protocol. The intermediate VL light chain library was harvested from the plates by mildly scraping the bacteria into 10 ml 2TY medium per plate. The cell mass was determined by OD600 measurement and two times 500 OD of bacteria was used for maxi plasmid DNA preparation using two P500 maxiprep columns (Macherey Nagel) according to manufacturer's instructions.

Analogous to the VL variable regions, the second round VH-JH products were first mixed together to obtain the normal J segment usage distribution (see Table 4), resulting in 7 VH subpools called PH1 to PH7. The pools were mixed to acquire a normalized sequence distribution using the percentages depicted in Table 4, obtaining one VH fraction that was digested with SfiI and XhoI restriction enzymes and ligated in SfiI-XhoI cut PDV-VL intermediate library obtained as described above. The ligation set- up, purification method, subsequent transformation of TG1 and harvest of bacteria was essentially as described for the VL intermediate library (see above) with the exception that 20 transformations and 20 square petridishes were used. The final library (approximately 1x10 cfu) was checked for insert frequency with a colony PCR using a primer set flanking the inserted VH-VL regions. 90% of the colonies showed a correct length insert. The colony PCR products were used for subsequent DNA sequence analysis to check sequence variation and to assess the percentage of colonies showing a complete ORF. This was 76% . Finally, the library was rescued and amplified by using CT helper phages (see WO 02/103012) and was used for phage antibody selection by panning methods as described below.

Example 2 Selection of phages carrying single chain Fv fragments against Influenza A and Influenza B heamagglutinin Antibody fragments were selected using antibody phage display libraries constructed essentially as described above and general phage display technology and MABSTRACT technology essentially as described in US Patent Number 6,265,150 and in WO 98/15833 (both of which are incorporated by reference herein). Furthermore, the methods and helper phages as described in WO 02/103012 (which is incorporated by reference herein) were used in the present invention.

Selection was performed against recombinant hemagglutinin (HA) of influenza A subtype H1 (A/New Caledonia/20/99), H3 (A/Wisconsin/67/2005), H4 (A/Duck/Hong Kong/24/1976), H5 (A/Chicken/Vietnam/28/2003), H7 (A/Netherlands/219/2003) and H9 (A/HongKong/1073/99). HA antigens were diluted in PBS (5.0 μg/ml), added to MaxiSorp Nunc-Immuno Tubes (Nunc) and incubated overnight at 4 C on a rotating wheel. The immunotubes were emptied and washed three times in block buffer (2% non- fat dry milk (ELK) in PBS). Subsequently, the immunotubes were filled completely with block buffer and incubated for 1-2 hrs at room temperature. Aliquots of phage display 13 13 library (500-1000 µl, 0.5x10 – 1x10 cfu, amplified using CT helper phage (see WO 02/103012)) were blocked in blocking buffer supplemented with 10% non-heat inactivated fetal bovine serum and 2% mouse serum for 1-2 hrs at room temperature. The blocked phage library was added to the immunotubes, incubated for 2 hrs at room temperature, and washed with wash buffer (0.05% (v/v) Tween-20 in PBS) to remove unbound phages. Bound phages were eluted from the respective antigen by incubation with 1 ml of 100 mM triethylamine (TEA) for 10 min at room temperature. Subsequently, the eluted phages were mixed with 0.5 ml of 1 M Tris-HCl pH 7.5 to neutralize the pH.

This mixture was used to infect 5 ml of an XL1-Blue E.coli culture that had been grown at 37°C to an OD 600 nm of approximately 0.3. The phages were allowed to infect the XL1-Blue bacteria for 30 min at 37°C. Then, the mixture was centrifuged for 10 min at 3000xg at room temperature and the bacterial pellet was resuspended in 0.5 ml 2-trypton yeast extract (2TY) medium. The obtained bacterial suspension was divided over two 2TY agar plates supplemented with tetracycline, ampicillin and glucose. After incubation overnight of the plates at 37ºC, the colonies were scraped from the plates and used to prepare an enriched phage library, essentially as described by De Kruif et al. (1995) and WO 02/103012. Briefly, scraped bacteria were used to inoculate 2TY medium containing ampicillin, tetracycline and glucose and grown at a temperature of 37ºC to an OD 600 nm of ~0.3. CT helper phages were added and allowed to infect the bacteria after which the medium was changed to 2TY containing ampicillin, tetracycline and kanamycin.

Incubation was continued overnight at 30ºC. The next day, the bacteria were removed from the 2TY medium by centrifugation after which the phages in the medium were precipitated using polyethylene glycol (PEG) 6000/NaCl. Finally, the phages were dissolved in 2 ml of PBS with 1% bovine serum albumin (BSA), filter-sterilized and used for the next round of selection. The second round of selection is performed either on the same HA subtype and/or on HA of a different subtype.

Two consecutive rounds of selections were performed before isolation of individual single-chain phage antibodies. After the second round of selection, individual E.coli colonies were used to prepare monoclonal phage antibodies. Essentially, individual colonies were grown to log-phase in 96 well plate format and infected with VCS-M13 helper phages after which phage antibody production was allowed to proceed overnight.

Phagemids were sequence analysed and all unique phagemids were used for further analysis. The supernatants containing phage antibodies were used directly in ELISA for binding to HA antigens. Alternatively, phage antibodies were PEG/NaCl-precipitated and filter-sterilized for both elisa and flow cytometry analysis.

Example 3 Validation of the HA specific single-chain phage antibodies Selected supernatants containing single-chain phage antibodies that were obtained in the screenings described above were validated in ELISA for specificity, i.e. binding to different HA antigens. For this purpose, baculovirus expressed recombinant H1 (A/New Caledonia/20/99), H3 (A/Wisconsin/67/2005), H5 (A/Vietnam/1203/04) H7 (A/Netherlands/219/2003), and B (B/Ohio/01/2005) HAs (Protein Sciences, CT, USA) were coated to Maxisorp ELISA plates. After coating, the plates were washed three times with PBS containing 0.1% v/v Tween-20 and blocked in PBS containing 3% BSA or 2% ELK for 1 hr at room temperature. The selected single-chain phage antibodies were incubated for 1 hr in an equal volume of PBS containing 4% ELK to obtain blocked phage antibodies. The plates were emptied, washed three times with PBS/0.1% Tween-20 and the blocked single-chain phage antibodies were added to the wells. Incubation was allowed to proceed for one hr, the plates were washed with PBS/0.1% Tween-20 and bound phage antibodies were detected (using OD 492nm measurement) using an anti- M13 antibody conjugated to peroxidase. As a control, the procedure was performed simultaneously without single-chain phage antibody and with an unrelated negative control single-chain phage antibody. From the selections on the different HA antigens with the phage libraries, 13 unique single-chain phage antibodies specifically binding recombinant influenza A H1, H3, H5, H7 and influenza B HA were obtained (SC09-003, SC09-004, SC09-005, SC09-006, SC09-007, SC09-008, SC09-009, SC09-010, SC09- 011, SC09-030, SC09-112, SC09-113 and SC09-114). See Table 5.

Alternatively, PEG/NaCl-precipitated and filter-sterilized phage antibodies were used to validate binding and specificity by FACS analysis. For this purpose, full-length recombinant influenza A subtypes H1 (A/New Caledonia/20/1999), H3 (A/Wisonsin/67/2005) and H7 (A/Netherlands/219/2003) HAs were expressed on the surface of PER.C6 cells. The cells were incubated with single-chain phage antibodies for 1 hr followed by three wash steps with PBS+0.1%BSA. Bound phages were detected using FITC conjugated M13-antibody. From the selections on the different HA antigens with the phage libraries, 14 single-chain phage antibodies specifically binding influenza A subtypes H1, H3 and H7 HA were found (SC09-003, SC09-004, SC09-005, SC09-006, SC09-007, SC09-008, SC09-009. SC09-010, SC09-011, SC09-012, SC09-030, SC09- 112, SC09-113 and SC09-114). See table 6.

All 16 phage antibodies, SC09-003, SC09-004, SC09-005, SC09-006, SC09-007, SC09-008, SC09-009. SC09-010, SC09-011, SC09-012, SC09-029, SC09-030, SC09- 031, SC09-112, SC09-113 and SC09-114, were used for construction of fully human immunoglobulins.

Example 4 Construction of fully human immunoglobulin molecules (human monoclonal antibodies) from the selected single chain Fvs From the selected specific single-chain phage antibodies (scFv) clones plasmid DNA was obtained and nucleotide and amino acid sequences were determined according to standard techniques. Heavy and light chain variable regions of the scFvs were cloned directly by restriction digest for expression in the IgG expression vectors pIg-C911- HCgamma1 (see SEQ ID N0: 175), pIG-C909-Ckappa (see SEQ ID NO: 176), or pIg- C910-Clambda (see SEQ ID No: 177). The VH and VL gene identity (see Tomlinson IM et al. V-BASE Sequence Directory. Cambridge United Kingdom: MRC Centre for Protein Engineering (1997)) of the scFvs were determined (see Table 7).

Nucleotide sequences for all constructs were verified according to standard techniques known to the skilled artisan. The resulting expression constructs encoding the human IgG1 heavy and light chains were transiently expressed in combination in 293T cells and supernatants containing human IgG1 antibodies were obtained and produced using standard purification procedures.

The amino acid sequence of the CDRs of the heavy and light chains of the selected immunoglobulin molecules is given in Table 7.

The number of amino-acid differences and the % identity of all heavy and light chain variable domains is given in Table 8.

Example 5 Cross-binding reactivity of IgGs A panel of five of the IgG antibodies described above, CR9005, CR9030, CR9112, CR9113 and CR9114, was validated in ELISA for binding specificity, i.e. binding to different HA antigens. For this purpose, baculovirus expressed recombinant H1 (A/New Caledonia/20/1999), H3 (A/Wisconsin/67/2005), H5 (A/Vietnam/1203/04, H7 (A/Netherlands/219/2003) and H9 (A/HongKong/1073/99) HA’s (Protein Sciences, CT, USA) were coated to Maxisorp ELISA plates. After coating, the plates were washed three times with PBS containing 0.1% v/v Tween-20 and blocked in PBS containing 3% BSA or 2% ELK for 1 hr at room temperature. The plates were emptied, washed three times with PBS/0.1% Tween-20 and the IgG antibodies were added to the wells. Incubation was allowed to proceed for one hr, the plates were washed with PBS/0.1% Tween-20 and bound antibodies were detected (using OD 492nm measurement) using an anti-human IgG antibody conjugated to peroxidase. As a control, an unrelated IgG CR4098 was used.

CR9005, CR9030, CR9112, CR9113 and CR9114 were shown to have heterosubtypic cross-binding activity to all the recombinant HAs tested. See table 9.

Additionally, the selected antibodies were used to test heterosubtypic binding by FACS analysis. For this purpose, full-length recombinant influenza A subtypes H1 (A/New Caledonia/20/1999), H3 (A/Wisonsin/67/2005) and H7 (A/Netherlands/219/2003) HAs were expressed on the surface of PER.C6 cells. The cells were incubated with IgG antibodies for 1 hr followed by three wash steps with PBS+0.1%BSA. Bound antibodies were detected using PE conjugated anti-human antibody. As a control, untransfected PER.C6 cells were used. CR9005, CR9030, CR9112, CR9113 and CR9114 show cross-binding activity to influenza A subtypes H1, H3 and H7 HA but not wild-type PER.C6 cells. See table 9.

Example 6 Cross-neutralizing activity of IgGs In order to determine whether the selected IgGs were capable of blocking multiple influenza A strains, additional in vitro virus neutralization assays (VNA) were performed.

The VNA were performed on MDCK cells (ATCC CCL-34). MDCK cells were cultured in MDCK cell culture medium (MEM medium supplemented with antibiotics, 20 mM Hepes and 0.15% (w/v) sodium bicarbonate (complete MEM medium), supplemented with 10% (v/v) fetal bovine serum). The H1 (A/WSN/33, A/New Caledonia/20/1999, A/Solomon Islands/IVR-145 (high-growth reassortant of A/Solomon Islands/3/2006), A/Brisbane/59/2007, A/NYMC/X-181 (high-growth reassortant of A/California/07/2009), H2 (A/Env/MPU3156/05), H3 (A/Hong Kong/1/68, A/Johannesburg/33/94, A/Panama/2000/1999, A/Hiroshima/52/2005, A/Wisconsin/67/2005 and A/Brisbane/10/2007), H4 (A/WF/HK/MPA892/06), H5 (PR8- H5N1-HK97 (6:2 reassortant of A/Hong Kong/156/97 and A/PR/8/34) and A/Eurasian Wigeon/MPF461/07), H6 (A/Eurasian Wigeon/MPD411/07), H7 (NIBRG-60 (6:2 reassortant of A/Mallard/Netherlands/12/2000) and PR8-H7N7-NY (7:1 reassortant of A/New York/107/2003 (H7N7) and A/PR/8/34)), H8 (A/Eurasian Wigeon/MPH571/08) H9 (A/Hong Kong/1073/99 and A/Chick/HK/SSP176/09), H10 (A/Chick/Germany/N/49) and H14 (PR8-H14N5 (6:2 reassortant of A/mallard/Astrakhan/263/1982 (H14N5) and A/PR/8/34)) strains which were used in the assay were all diluted to a titer of 5,7 x10 TCID50/ml (50% tissue culture infective dose per ml), with the titer calculated according to the method of Spearman and Karber. The IgG preparations (200 μg/ml) were serially 2-fold diluted (1:2 - 1:512) in complete MEM medium in quadruplicate wells. 25 μl of the respective IgG dilution was mixed with 25 μl of virus suspension (100 TCID50/25 μl) and incubated for one hr at 37°C. The suspension was then transferred in quadruplicate onto 96-well plates containing confluent MDCK cultures in 50 μl complete MEM medium. Prior to use, MDCK cells were seeded at 3x10 cells per well in MDCK cell culture medium, grown until cells had reached confluence, washed with 300-350 μl PBS, pH 7.4 and finally 50 μl complete MEM medium was added to each well. The inoculated cells were cultured for 3-4 days at 37°C and observed daily for the development of cytopathogenic effect (CPE). CPE was compared to the positive control.

CR9005, CR9112, CR9113 and CR9114 show heterosubtypic cross-neutralizing activity to representative strains of all tested influenza A subtypes H1, H2, H3, H4, H5, H6, H7, H8, H9 and H10 viruses. See table 10.

Example 7 Pan-influenza antibodies bind to the pre-fusion conformation of HA In order to determine whether the selected IgGs were capable of binding the pre- or post-fusion conformation of the HA molecule, an in vitro pH-shift experiment was performed. For this purpose, full-length recombinant influenza A subtypes H1 (A/New Caledonia/20/99), H3 (A/Wisonsin/67/2005), H5 (A/Vietnam/1203/04), H7 (A/Netherlands/219/03) and H9 (A/Hong Kong/1073/99) HA were expressed on the surface of PER.C6 cells. To measure mAb binding to different structural HA conformations, cells were detached from the plastic support using PBS-EDTA and subsequently treated with trypsin (TrypLE Select, Gibco) for 5 min at RT, washed (1% BSA in PBS) and incubated for 15 min in citric acid–sodium phosphate buffer (pH 4.9).

Cell samples were set aside after each processing step (untrypsinized/HA0; trypsinized/HA1-HA2; pH 4.9/fusion HA) and fractions of each treatment were incubated with mAb CR9114 for 1 hour. Cells were then incubated for 30 min with phycoerythrin- conjugated anti-human IgG (Southern Biotech) in 1% BSA. Stained cells were analysed using a FACS Canto with FACS Diva software (Becton Dickinson).

FACS binding of IgG1s to surface expressed HA was after sequential treatment with trypsin and pH 4.9 buffered medium and expressed as percentage binding to untreated HA (A). See Figure 1A.

Antibody CR9114 shows a marked decrease in binding after pH-shift indicating specificity for an epitope present only before the low pH induced conformational change of the HA molecule.

Alternatively, to test whether the IgGs can block the low pH induced conformational change of HA, antibody CR9114 was added before the low pH step.

Samples of consecutive treatments were split and stained with either phycoerythrin- conjugated anti-human IgG (Southern Biotech). Stained cells were analysed using a FACS Canto with FACS Diva software (Becton Dickinson). See Fig 1B.

Antibody CR9114 shows a high level of residual binding to the various HAs after pH- shift indicating that when these antibodies are bound to the HA molecule, the low pH induced conformational change does not occur.

Example 8 Affinity measurements of Fabs on various influenza A and B HAs.

Recombinant soluble HA of A/New Caledonia/20/1999 (H1), A/Brisbane/59/2007 (H1), A/Wisconsin/67/2005 (H3), A/Brisbane/10/2007 (H3, B/Florida/4/2006 (B), B/Brisbane/60/2008 (B) and B/Malaysia/2506/2004 (B) produced using baculovirus vectors in insect cells were purchased from Protein Sciences Corp (CT, USA) and biotinylated at room temperature (RT) for 40 min using EZ-link sulfo-NHS-LC-LC- biotin (Pierce). Buffer exchange step to PBS was performed using Amicon Ultra 0.5 ml Centrifugal Filters (Millipore). Biotinylated HA was bound to Streptavidin sensors at 37 °C for 1200 seconds. Association of Fab fragment of CR9005, CR9112, CR9113 and CR9114 to HA was measured on Octet QK (ForteBio) for 700 seconds at 37 °C by exposing the sensors to 100 nM antibody in 1x kinetic buffer (ForteBio). Dissociation of the Fab fragments was assessed by exposing the sensors to 1x kinetic buffer for 9000 seconds at 37 °C. Fab fragments of CR9005, CR9112, CR9113 and CR9114 all bind with micro- to pico-molar affinities to H1, H3 and influenza B HA.

Example 9 Competition for binding with other stem binding antibodies Recombinant soluble HA of A/New Caledonia/20/1999 (H1N1) and A/Wisconsin/67/2005 (H3N2) produced using baculovirus vectors in insect cells were purchased from Protein Sciences Corp (CT, USA) and biotinylated at room temperature (RT) for 40 min using EZ-link sulfo-NHS-LC-LC-biotin (Pierce). Buffer exchange step to PBS was performed using Amicon Ultra 0.5 ml Centrifugal Filters (Millipore).

Biotinylated HA was bound to Streptavidin sensors at 37 °C for 1200 seconds.

Association of antibodies CR9114 and CR6261 to H1 HA was measured on Octet QK (ForteBio) for 700 seconds at 37 °C by exposing the sensors to 100 nM antibody in 1x kinetic buffer (ForteBio) after which the degree of additional binding was assessed by exposing the sensors to a second antibody (100 nM in 1x kinetic buffer) in the presence of the first antibody (100 nM) for 700 seconds at 37 °C. As a control, mAb CR9020, binding to the globular head of H1 was taken along. Association of antibodies CR9114 and CR8020 to H3 HA was measured on Octet QK (ForteBio) for 900 seconds at 37 °C by exposing the sensors to 100 nM antibody in 1x kinetic buffer (ForteBio) after which the degree of additional binding was assessed by exposing the sensors to a second antibody (100 nM in 1x kinetic buffer) in the presence of the first antibody (100 nM) for 900 seconds at 37 °C. As a control, mAb CR8057, binding to the globular head of H3 was taken along.

CR9114 competes for binding to H1 HA with CR6261 and to H3 HA with CR8020. CR9114 therefore likely binds an epitope overlapping with both the epitopes of CR6261 and CR8020 in the stem-region of HA. (See Fig. 2) Example 10 Prophylactic activity of human IgG monoclonal antibody CR9114 against lethal influenza B challenge in vivo A study was performed to test the prophylactic effect of the monoclonal antibody CR9114 against a lethal challenge with influenza B virus in vivo. MAb CR9114 was tested for prophylactic efficacy in a mouse lethal challenge model with mouse adapted influenza B/Florida/04/2006 virus (Central Veterinary Institute (CVI), Lelystad, The Netherlands). The B/Florida/04/2006 virus was adapted to mice after 5 lung-to-lung passages. The mouse adapted influenza B passage 5 virus was propagated in embryonated chicken eggs in CVI’s laboratory. All mice (Balb/c, female, age 6-8 weeks, n=10 per group) were acclimatized and maintained for a period of at least 4 days prior to the start of the experiment. MAb CR9114 was dosed at 15 mg/kg intravenously in the tail vein (vena coccygeus) at day -1 before challenge, assuming an average weight of 18 g per mouse and a fixed dose volume of 0.2 mL. A control group was taken along dosed with vehicle control. The mice were then challenged at day 0 with 25 LD B/Florida/04/2006 influenza B virus by intranasal inoculation. Clinical signs and body weights were determined daily from day-1 before challenge until day 8. Clinical signs were scored with a scoring system (0=no clinical signs; 1=rough coat; 2=rough coat, less reactive during handling; 3=rough coat, rolled up, laboured breathing, less reactive during handling; 4=rough coat, rolled up, laboured breathing, inactive response to manipulation/handlings). At a score of 4 the animal was euthanized.

All mice were active and appeared healthy without showing signs of disease during the acclimatization period. Fig. 3A shows the survival rates of the mice, following mAb administration. Mice dosed with 15 mg/kg mAb CR9114 showed a survival rate of 100%, whereas in the control mAb group 50% survived.

In Figure 3B the mean body weight change of the mice during the 8 day study period following mAb administration is shown. In the mAb CR9114 group the mice did not loose weight over the 8 day study period, whereas in the vehicle control group weight loss was observed. Median clinical scores of the mice are depicted in Fig. 3C. Of the mice treated with 15 mg/kg mAb CR9114 at day -1 pre-challenge, all survived and none of the animals showed any clinical signs during the observation period (from day 0 to day 8 post infection). These results show that the human anti influenza antibody CR9114, identified and developed as disclosed herein, is able to provide protection against a lethal dose of influenza B virus in vivo. When administered one day prior to infection at a dose of 15 mg/kg or higher, mAb CR9114 was able to completely prevent clinical manifestation of influenza B infection in mice.

Table 1. First round Vkappa, Vlambda and VH amplifications Primer name Primer nucleotide sequence SEQ ID NO: OK1 (HuVK1B) GAC ATC CAG WTG ACC CAG TCT CC 65 OK2 (HuVK2) GAT GTT GTG ATG ACT CAG TCT CC 66 OK3 (HuVK2B2) GAT ATT GTG ATG ACC CAG ACT CC 67 OK4 (HuVK3B) GAA ATT GTG WTG ACR CAG TCT CC 68 OK5 (HuVK5) GAA ACG ACA CTC ACG CAG TCT CC 69 OK6 (HuVK6) GAA ATT GTG CTG ACT CAG TCT CC 70 OCK (HuCK) ACA CTC TCC CCT GTT GAA GCT CTT 71 OL1 (HuVL1A)* CAG TCT GTG CTG ACT CAG CCA CC 72 OL1 (HuVL1B)* CAG TCT GTG YTG ACG CAG CCG CC 73 OL1 (HuVL1C)* CAG TCT GTC GTG ACG CAG CCG CC 74 OL2 (HuVL2B) CAG TCT GCC CTG ACT CAG CC 75 OL3 (HuVL3A) TCC TAT GWG CTG ACT CAG CCA CC 76 OL4 (HuVL3B) TCT TCT GAG CTG ACT CAG GAC CC 77 OL5 (HuVL4B) CAG CYT GTG CTG ACT CAA TC 78 OL6 (HuVL5) CAG GCT GTG CTG ACT CAG CCG TC 79 OL7 (HuVL6) AAT TTT ATG CTG ACT CAG CCC CA 80 OL8 (HuVL7/8) CAG RCT GTG GTG ACY CAG GAG CC 81 OL9 (HuVL9)# CWG CCT GTG CTG ACT CAG CCM CC 82 OL9 (HuVL10)# CAG GCA GGG CTG ACT CAG 83 OCL (HuCL2)X TGA ACA TTC TGT AGG GGC CAC TG 84 OCL (HuCL7)X AGA GCA TTC TGC AGG GGC CAC TG 85 OH1(HuVH1B7A)+ CAG RTG CAG CTG GTG CAR TCT GG 86 OH1 (HuVH1C)+ SAG GTC CAG CTG GTR CAG TCT GG 87 OH2 (HuVH2B) CAG RTC ACC TTG AAG GAG TCT GG 88 OH3 (HuVH3A) GAG GTG CAG CTG GTG GAG 89 OH4 (HuVH3C) GAG GTG CAG CTG GTG GAG WCY GG 90 OH5 (HuVH4B) CAG GTG CAG CTA CAG CAG TGG GG 91 OH6 (HuVH4C) CAG STG CAG CTG CAG GAG TCS GG 92 OH7 (HuVH6A) CAG GTA CAG CTG CAG CAG TCA GG 93 OCM (HuCIgM) TGG AAG AGG CAC GTT CTT TTC TTT 94 * Mix in 1:1:1 ratio # Mix in 1:1 ratio X Mix in 1:1 ratio + Mix in 1:1 ratio Table 2. Second round Vkappa, Vlambda and VH amplifications Primer name Primer nucleotide sequence SEQ ID NO OK1S (HuVK1B-SAL) TGA GCA CAC AGG TCG ACG GAC ATC CAG 95 WTG ACC CAG TCT CC OK2S (HuVK2-SAL) TGA GCA CAC AGG TCG ACG GAT GTT GTG 96 ATG ACT CAG TCT CC OK3S (HuVK2B2-SAL) TGA GCA CAC AGG TCG ACG GAT ATT GTG 97 ATG ACC CAG ACT CC OK4S (HuVK3B-SAL) TGA GCA CAC AGG TCG ACG GAA ATT GTG 98 WTG ACR CAG TCT CC OK5S (HuVK5-SAL) TGA GCA CAC AGG TCG ACG GAA ACG ACA 99 CTC ACG CAG TCT CC OK6S (HuVK6-SAL) TGA GCA CAC AGG TCG ACG GAA ATT GTG 100 CTG ACT CAG TCT CC OJK1 (HuJK1-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACG 101 TTT GAT TTC CAC CTT GGT CCC OJK2 (HuJK2-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACG 102 TTT GAT CTC CAG CTT GGT CCC OJK3 (HuJK3-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACG 103 TTT GAT ATC CAC TTT GGT CCC OJK4 (HuJK4-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACG 104 TTT GAT CTC CAC CTT GGT CCC OJK5 (HuJK5-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACG 105 TTT AAT CTC CAG TCG TGT CCC OL1S (HuVL1A-SAL)* TGA GCA CAC AGG TCG ACG CAG TCT GTG 106 CTG ACT CAG CCA CC OL1S (HuVL1B-SAL)* TGA GCA CAC AGG TCG ACG CAG TCT GTG 107 YTG ACG CAG CCG CC OL1S (HuVL1C-SAL)* TGA GCA CAC AGG TCG ACG CAG TCT GTC 108 GTG ACG CAG CCG CC OL2S (HuVL2B-SAL) TGA GCA CAC AGG TCG ACG CAG TCT GCC 109 CTG ACT CAG CC OL3S (HuVL3A-SAL) TGA GCA CAC AGG TCG ACG TCC TAT GWG 110 CTG ACT CAG CCA CC OL4S (HuVL3B-SAL) TGA GCA CAC AGG TCG ACG TCT TCT GAG 111 CTG ACT CAG GAC CC OL5S (HuVL4B-SAL) TGA GCA CAC AGG TCG ACG CAG CYT GTG 112 CTG ACT CAA TC OL6S (HuVL5-SAL) TGA GCA CAC AGG TCG ACG CAG GCT GTG 113 CTG ACT CAG CCG TC OL7S (HuVL6-SAL) TGA GCA CAC AGG TCG ACG AAT TTT ATG 114 CTG ACT CAG CCC CA OL8S (HuVL7/8-SAL) TGA GCA CAC AGG TCG ACG CAG RCT GTG 115 GTG ACY CAG GAG CC OL9S (HuVL9-SAL)# TGA GCA CAC AGG TCG ACG CWG CCT GTG 116 CTG ACT CAG CCM CC OL9S (HuVL10-SAL)# TGA GCA CAC AGG TCG ACG CAG GCA GGG 117 CTG ACT CAG OJL1 (HuJL1-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACC 118 TAG GAC GGT GAC CTT GGT CCC OJL2 (HuJL2/3-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACC 119 TAG GAC GGT CAG CTT GGT CCC OJL3 (HuJL7-NOT) GAG TCA TTC TCG ACT TGC GGC CGC ACC 120 GAG GAC GGT CAG CTG GGT GCC OH1S (HuVH1B-SFI)+ GTC CTC GCA ACT GCG GCC CAG CCG GCC 121 ATG GCC CAG RTG CAG CTG GTG CAR TCT GG OH1S (HuVH1C-SFI)+ GTC CTC GCA ACT GCG GCC CAG CCG GCC 122 ATG GCC SAG GTC CAG CTG GTR CAG TCT GG OH2S (HuVH2B-SFI) GTC CTC GCA ACT GCG GCC CAG CCG GCC 123 ATG GCC CAG RTC ACC TTG AAG GAG TCT GG OH3S (HuVH3A-SFI) GTC CTC GCA ACT GCG GCC CAG CCG GCC 124 ATG GCC GAG GTG CAG CTG GTG GAG OH4S (HuVH3C-SFI) GTC CTC GCA ACT GCG GCC CAG CCG GCC 125 ATG GCC GAG GTG CAG CTG GTG GAG WCY GG OH5S (HuVH4B-SFI) GTC CTC GCA ACT GCG GCC CAG CCG GCC 126 ATG GCC CAG GTG CAG CTA CAG CAG TGG GG OH6S (HuVH4C-SFI) GTC CTC GCA ACT GCG GCC CAG CCG GCC 127 ATG GCC CAG STG CAG CTG CAG GAG TCS GG OH7S (HuVH6A-SFI) GTC CTC GCA ACT GCG GCC CAG CCG GCC 128 ATG GCC CAG GTA CAG CTG CAG CAG TCA GG OJH1 (HuJH1/2-XHO) GAG TCA TTC TCG ACT CGA GAC RGT GAC 129 CAG GGT GCC OJH2 (HuJH3-XHO) GAG TCA TTC TCG ACT CGA GAC GGT GAC 130 CAT TGT CCC OJH3 (HuJH4/5-XHO) GAG TCA TTC TCG ACT CGA GAC GGT GAC 131 CAG GGT TCC OJH4 (HuJH6-XHO) GAG TCA TTC TCG ACT CGA GAC GGT GAC 132 CGT GGT CCC * Mix in 1:1:1 ratio # Mix in 1:1 ratio + Mix in 1:1 ratio Table 3. Second round VL regions amplification overview Template 5’ 3’ Product Share in Pool Share in primer primer PK/PL(%) VL (%) OK1S OJK1 K1J1 25 OK1S OJK2 K1J2 25 K1 OK1S OJK3 K1J3 10 PK1 30 OK1S OJK4 K1J4 25 OK1S OJK5 K1J5 15 OK2S OJK1 K2J1 25 OK2S OJK2 K2J2 25 K2 OK2S OJK3 K2J3 10 PK2 4 OK2S OJK4 K2J4 25 OK2S OJK5 K2J5 15 OK3S OJK1 K3J1 25 OK3S OJK2 K3J2 25 K3 OK3S OJK3 K3J3 10 PK3 1 OK3S OJK4 K3J4 25 OK3S OJK5 K3J5 15 OK4S OJK1 K4J1 25 OK4S OJK2 K4J2 25 K4 OK4S OJK3 K4J3 10 PK4 19 OK4S OJK4 K4J4 25 OK4S OJK5 K4J5 15 OK5S OJK1 K5J1 25 OK5S OJK2 K5J2 25 K5 OK5S OJK3 K5J3 10 PK5 1 OK5S OJK4 K5J4 25 OK5S OJK5 K5J5 15 OK6S OJK1 K6J1 25 OK6S OJK2 K6J2 25 K6 OK6S OJK3 K6J3 10 PK6 5 OK6S OJK4 K6J4 25 OK6S OJK5 K6J5 15 OL1S OJL1 L1J1 30 L1 OL1S OJL2 L1J2 60 PL1 14 OL1S OJL3 L1J3 10 OL2S OJL1 L2J1 30 L2 OL2S OJL2 L2J2 60 PL2 10 OL2S OJL3 L2J3 10 OL3S OJL1 L3J1 30 L3 OL3S OJL2 L3J2 60 PL3 10 OL3S OJL3 L3J3 10 OL4S OJL1 L4J1 30 L4 OL4S OJL2 L4J2 60 PL4 1 OL4S OJL3 L4J3 10 OL5S OJL1 L5J1 30 L5 OL5S OJL2 L5J2 60 PL5 1 OL5S OJL3 L5J3 10 OL6S OJL1 L6J1 30 L6 OL6S OJL2 L6J2 60 PL6 1 OL6S OJL3 L6J3 10 OL7S OJL1 L7J1 30 L7 OL7S OJL2 L7J2 60 PL7 1 OL7S OJL3 L7J3 10 OL8S OJL1 L8J1 30 L8 OL8S OJL2 L8J2 60 PL8 1 OL8S OJL3 L8J3 10 OL9S OJL1 L9J1 30 L9 OL9S OJL2 L9J2 60 PL9 1 OL9S OJL3 L9J3 10 VL 100% Table 4. Second round VH regions amplification overview Template 5’ 3’ Product Share in Pool Share in primer primer PK/PL VH (%) OH1S OJH1 H1J1 10 OH1S OJH2 H1J2 10 H1 OH1S OJH3 H1J3 60 PH1 25 OH1S OJH4 H1J4 20 OH2S OJH1 H2J1 10 OH2S OJH2 H2J2 10 H2 OH2S OJH3 H2J3 60 PH2 2 OH2S OJH4 H2J4 20 OH3S OJH1 H3J1 10 OH3S OJH2 H3J2 10 H3 OH3S OJH3 H3J3 60 PH3 25 OH3S OJH4 H3J4 20 OH4S OJH1 H4J1 10 OH4S OJH2 H4J2 10 H4 OH4S OJH3 H4J3 60 PH4 25 OH4S OJH4 H4J4 20 OH5S OJH1 H5J1 10 OH5S OJH2 H5J2 10 H5 OH5S OJH3 H5J3 60 PH5 2 OH5S OJH4 H5J4 20 OH6S OJH1 H6J1 10 OH6S OJH2 H6J2 10 H6 OH6S OJH3 H6J3 60 PH6 20 OH6S OJH4 H6J4 20 OH7S OJH1 H7J1 10 OH7S OJH2 H7J2 10 H7 OH7S OJH3 H7J3 60 PH7 1 OH7S OJH4 H7J4 20 VH 100% Table 5: Cross-binding activity of PEG/NACl-precipitated and filter-sterilized single- chain phage antibodies to HA of different subtypes, as measured by ELISA. + = binding (>4x background); +/- = low binding (2-4x background) - = no detectable binding; H1= HA of influenza A H1 subtype; H3= HA of influenza A H3 subtype; H5=HA of influenza A H5 subtype; H7=HA of influenza A H7 subtype; B= HA of influenza virus B; Rabies=Glycoprotein of Rabies virus (negative control).

Phage midi Elisa H1 H3 H5 H7 B Rabies sc09-003 + + + + + - sc09-004 + + + + + - sc09-005 + + + + + - sc09-006 + + + + + - sc09-007 + +/- + + +/- - sc09-008 + +/- + + +/- - sc09-009 + +/- + + +/- - sc09-010 + + + + +/- - sc09-011 + + + + + - sc09-012 + + + + - - sc09-029 + +/- + + - - sc09-030 + + + + + - sc09-031 + +/- + + - - sc09-112 + + + + + - sc09-113 + + + + + - sc09-114 + + + + + - Table 6. FACS analysis of PEG/NACl-precipitated and filter-sterilized phage antibodies. + = binding (>4x background); +/- = low binding (2-4x background) - = no detectable binding; PER.C6=untransfected PER.C6 cells (control); mH1, mH3, mH7= membrane bound HA of the subtypes H1, H3 and H7 subtypes, respectively.

Phage midi Facs (% gated UL) PerC6 mH1 mH3 mH7 sc09-003 - + + + sc09-004 - + + + sc09-005 - + + + sc09-006 - + + + sc09-007 - + +/- + sc09-008 - + +/- + sc09-009 - + +/- + sc09-010 - + + + sc09-011 - + + + sc09-012 - + + + sc09-029 - + - +/- sc09-030 - + + + sc09-031 - + - +/- sc09-112 - + + + sc09-113 - + + + sc09-114 - + + + Table 7. Data of the CDR regions of the HA specific immunoglobulins. The SEQ ID NO is given between brackets.

IgG# VH HC CDR1 HC CDR2 HC CDR3 VL LC CDR1 LC CDR2 LC CDR3 CR9003 IGHV1-69*06 GGTSNNFG (133) ISPIFGST (134) ARHGNYYFYSGMDL (135) IGLV3-21*02 NVGSNS (136) DDR (137) QVWDSSSDHRV (138) CR9004 IGHV1-69*06 GGTSNNYA (139) VSPIFGST (140) ARHGNYYYNSGMDV (141) IGLV1-44*01 DSNIGRRS (142) SND (143) AAWDDSLKGAV (144) CR9005 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDL (145) IGLV2-14*01 SSDVGGYNY (146) DVS (174) CSYAGSAKGV (147) CR9006 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDL (145) IGLV3-21*02 NIGSKT (148) GDS (149) QVWDSSSDHPGAV (150) CR9007 IGHV1-69*06 GGTSNNYA (139) ISPIFGSA (151) ARHGNYYYYSGMDV (152) IGLV1-44*01 SSNIGSNT (153) GDD (154) ATWDDSLNGHV (155) CR9008 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGLV3-21*02 NIGSKT (148) GDS (149) QVWDSSSDHPGAV (150) CR9009 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGKV1-12*01 QHISSW (156) SAS (157) QQANSFPLT (158) CR9010 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGLV3-21*02 NIGSKT (148) VDS (159) QVWDSNSDHPGAV (160) CR9011 IGHV1-69*06 GGTSNNYA (139) ISPIFGSA (151) ARHGNYYYYSGTDV (161) IGLV1-44*01 DSNIGRRS (142) SND (143) AAWDDSLKGAV (144) CR9012 IGHV1-69*06 GGTSNNYA (139) ISPIFGSA (151) ARHGTYYYYSGMDV (162) IGLV1-40*02 SSNIGAGYD (163) GNN (164) QSYDQNLSEGV (165) CR9029 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGKV3-20*01 QSVSSY (166) GAS (167) QQYGSSPFA (168) CR9030 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGLV3-21*02 NIGSKS (169) GDS (149) QVWDSSSDHPGAV (150) CR9031 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYNSGMDV (141) IGLV1-40*01 SSNIGAGYD (163) DNN (169) QSYDSGLSASPYV (170) CR9112 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGLV1-40*01 SANIGAGYD (171) GNN (164) QSYDSSLSGAL (172) CR9113 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDL (145) IGLV1-44*01 DSNIGRRS (142) SND (143) AAWDASLSGPV (173) CR9114 IGHV1-69*06 GGTSNNYA (139) ISPIFGST (134) ARHGNYYYYSGMDV (152) IGLV1-44*01 DSNIGRRS (142) SND (143) AAWDDSLKGAV (144) Table 8. Identity cross-tables of the amino acid sequences of the heavy and light chain variable domains.

Amino acid differences in Heavy Chain SC09-007 2 3 5 565455 6793 11 15 SC09-011 98.4 5 5 5 6 7 6 7 7 8 9 9 3 13 15 SC09-112 97.5 95.9 2 2 3 2 1 2 2 3 4 6 6 8 12 SC09-010 95.9 95.9 98.4 0 3 4 3 4 4 5 6 4 6 10 10 SC09-029 95.9 95.9 98.4 100.0 3 4 3 4 4 5 6 4 6 10 10 SC09-008 95.0 95.0 97.5 97.5 97.5 3 2 3 5 6 5 5 7 9 11 SC09-030 95.9 94.2 98.4 96.7 96.7 97.5 1 2 4 5 4 6 8 6 12 SC09-114 96.7 95.0 99.2 97.5 97.5 98.4 99.2 1 3 4 3 5 7 7 11 SC09-009 95.9 94.2 98.4 96.7 96.7 97.5 98.4 99.2 4 5 4 6 8 8 12 SC09-004 95.9 94.2 98.4 96.7 96.7 95.9 96.7 97.5 96.7 3 6 8 8 10 14 SC09-031 95.0 93.4 97.5 95.9 95.9 95.0 95.9 96.7 95.9 97.5 5 7 9 11 15 SC09-005 94.2 92.6 96.7 95.0 95.0 95.9 96.7 97.5 96.7 95.0 95.9 2 8 6 10 SC09-006 92.6 92.6 95.0 96.7 96.7 95.9 95.0 95.9 95.0 93.4 94.2 98.4 88 8 SC09-012 97.5 97.5 95.0 95.0 95.0 94.2 93.4 94.2 93.4 93.4 92.6 93.4 93.4 12 14 SC09-113 90.9 89.3 93.4 91.7 91.7 92.6 95.0 94.2 93.4 91.7 90.9 95.0 93.4 90.1 8 SC09-003 87.6 87.6 90.1 91.7 91.7 90.9 90.1 90.9 90.1 88.4 87.6 91.7 93.4 88.4 93.4 Percentage identity SC09-007 SC09-011 SC09-112 SC09-010 SC09-029 SC09-008 SC09-030 SC09-114 SC09-009 SC09-004 SC09-031 SC09-005 SC09-006 SC09-012 SC09-113 SC09-003 Amino acid differences in Light Chain SC09-011 0 2 7 14 29 26 34 44 47 47 45 52 47 62 64 SC09-114 100.0 2 7 14 29 26 34 44 47 47 45 52 47 62 64 SC09-004 98.2 98.2 5 16 27 24 32 42 49 49 47 54 49 62 64 SC09-113 93.6 93.6 95.5 17 25 22 29 41 46 46 44 51 47 62 64 SC09-007 87.3 87.3 85.5 84.6 26 25 32 42 41 41 41 47 43 61 61 SC09-012 73.9 73.9 75.7 77.5 76.6 9 13 39 48 48 47 52 48 61 62 SC09-112 76.6 76.6 78.4 80.2 77.5 91.9 13 37 45 45 44 51 45 60 60 SC09-031 69.9 69.9 71.7 74.3 71.7 88.5 88.5 37 50 50 49 53 46 60 62 SC09-005 60.4 60.4 62.2 63.1 62.2 64.9 66.7 67.3 55 55 54 56 46 64 63 SC09-006 58.0 58.0 56.3 58.9 63.4 57.5 60.2 55.8 51.3 0 3 7 17 64 61 SC09-008 58.0 58.0 56.3 58.9 63.4 57.5 60.2 55.8 51.3 100.0 3 7 17 64 61 SC09-030 59.8 59.8 58.0 60.7 63.4 58.4 61.1 56.6 52.2 97.3 97.3 10 14 62 59 SC09-010 53.6 53.6 51.8 54.5 58.0 54.0 54.9 53.1 50.4 93.6 93.6 90.9 22 67 67 SC09-003 57.7 57.7 55.9 57.7 61.3 57.1 59.8 59.3 58.6 84.6 84.6 87.3 80.0 62 56 SC09-009 45.1 45.1 45.1 45.1 46.0 46.5 47.4 47.4 43.4 42.9 42.9 44.6 40.2 44.1 34 SC09-029 43.4 43.4 43.4 43.4 46.0 45.6 47.4 45.6 44.3 45.5 45.5 47.3 40.2 49.6 68.2 Percentage identity SC09-011 SC09-114 SC09-004 SC09-113 SC09-007 SC09-012 SC09-112 SC09-031 SC09-005 SC09-006 SC09-008 SC09-030 SC09-010 SC09-003 SC09-009 SC09-029 Table 9. Cross-binding reactivity of IgGs, as measured by ELISA and FACS.

H1=soluble recombinant A/New Caledonia/20/1999 H1 HA; H3= soluble recombinant A/Wisconsin/67/2005 H3 HA; H5= soluble recombinant A/Vietnam/1203/04 H5 HA; H7= soluble recombinant A/Netherlands/219/2003 H7 HA; H9= soluble recombinant A/Hong Kong/1073/99 H9 HA; B= soluble recombinant B/Ohio/01/05 influenza B HA; Rabies= rabies glycoprotein; PER.C6=untransfected PER.C6 cells (control); mH1=PER.C6 expressed A/New Caledonia/20/1999 H1 HA; mH3= PER.C6 expressed A/Wisconsin/67/2005 H3 HA; mH7= PER.C6 expressed A/Netherlands/219/2003 H7 HA; ND=not done. + = binding (>10x background); +/- = low binding (2-10x background) - = no detectable binding.

IgG Elisa IgG Facs H1 H3 H5 H7 H9 B Rabies PerC6 mH1 mH3 mH7 CR9005 + + + + + + - - + + + CR9030 + + + + + +/- - - + + + CR9112 + + + + + + - - + + + CR9113 + + + + + + - - + + + CR9114 + + + + + + - - + + + CR4098 - - - - - - + - - - - Table 10. Cross-neutralizing activity of IgGs; Titers (indicated in µg/ml) are geomean IC50 values as determined according to the Spearman-Karber method of at least duplicate experiments; >100 = not neutralizing at highest tested concentration (100 µg/ml).

Subtype Strain CR9005 CR9112 CR9113 CR9114 Group I H1 A/WSN/33 1.1 0.9 1.1 1.1 A/New Caledonia/20/99 2.6 1.9 4.4 3.7 A/Solomon Islands/3/2006 1.4 1.3 2.2 1.8 A/Brisbane/59/2007 3.4 2 3.1 2.6 A/California/7/2009 0.7 0.5 0.3 0.3 H2 A/Env/MPU3156/05 8.8 6.3 8.8 8.8 H5 A/Hong Kong/156/97 0.8 0.7 0.9 0.4 A/EW/MPF461/07 10.5 10.5 8.8 10.5 H6 A/EW/MPD411/07 29.7 10.5 17.7 10.5 H8 A/EW/MPH571/08 8.8 8.8 8.8 8.8 H9 A/Hong Kong/1073/99 6.3 3.7 3.7 4.4 A/Ck/HK/SSP176/09 4.4 4.4 6.3 6.3 Group II H3 A/Hong Kong/1/68 42 27.6 22.3 19 A/Johannesburg/33/94 17.7 13.8 32.4 21.9 A/Panama/2007/1999 28.2 47.5 47.5 39.9 A/Hiroshima/52/2005 22.9 10.5 13.6 12.5 A/Wisconsin/67/2005 35.4 29.7 35.4 32.4 A/Brisbane/10/2007 11.2 5.6 9.4 5.6 H4 A/WF/MPA 892/06 1.2 0.8 1.3 0.8 H7 A/Mallard/Netherlands/12/2000 9.6 6.3 6.3 4.8 A/New York/107/2003 > 100 > 100 > 100 > 100 H10 A/Chick/Germany/N/49 29.6 26.5 19.8 15.7 H14 A/Mallard/Astrakhan/263/1982 > 100 > 100 > 100 > 100 SEQUENCES >SC09-003 VH DNA (SEQ ID NO: 1) GAGGTGCAGCTGGTGGAGTCTGGGGCTGAGGTCAAGAAGGCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAACAACTTTGGTATCAGCTGGGTACGACAGGCCCCTGGCCAAGGCCTTGAGTGGA TGGGCGGGATCAGCCCAATCTTTGGTTCGACAGTCTACGCACAGAAATTTCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCACACACTGCCTACATGGAGATGAACAGCCTGACATCTGAGGACACGGCCGTCTATTT CTGTGCGAGGCACGGAAATTATTATTTCTACTCCGGTATGGACCTCTGGGGCCAAGGGACCACGGTCACC >SC09-003 VH PROTEIN (SEQ ID NO: 2) EVQLVESGAEVKKAGSSVKVSCKSSGGTSNNFGISWVRQAPGQGLEWMGGISPIFGSTVYAQKFQGRVTIS ADIFSHTAYMEMNSLTSEDTAVYFCARHGNYYFYSGMDLWGQGTTVT >SC09-003 VL DNA (SEQ ID NO: 3) TCCTATGTGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGACAGACGGCCACGATTTCCTGTGGGGG AGACAACGTTGGAAGTAACAGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTGGTCGTCT ATGATGATCGCGACCGACCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGAACACGGCCACC CTGACCATCAGCAGGGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATAGTAGTAGTGA TCATCGAGTCTTCGGAACTGGGACCAAGGTCACCGTCCTAG >SC09-003 VL PROTEIN (SEQ ID NO: 4) SYVLTQPPSVSVAPGQTATISCGGDNVGSNSVHWYQQKPGQAPVLVVYDDRDRPSGIPERFSGSNSGNTAT LTISRVEAGDEADYYCQVWDSSSDHRVFGTGTKVTVL >SC09-004 VH DNA (SEQ ID NO: 5) CAGGTCCAGCTGGTACAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGCGGCACCTCCAATAACTATGCCATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGGTCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGTCTGACATCTGAGGACACGGCCGTCTATTA TTGTGCGAGACACGGGAATTATTATTACAACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACC >SC09-004 VH PROTEIN (SEQ ID NO: 6) QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGVSPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYYCARHGNYYYNSGMDVWGQGTTVT >SC09-004 VL DNA (SEQ ID NO: 7) CAGTCTGTGCTGACGCAGCCGCCCGCAGTGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCGTGTTCTGG AAGTGATTCCAACATCGGGAGAAGAAGTGTAAACTGGTACCAGCAGTTCCCAGGAACGGCCCCCAAACTCC TCATCTATAGTAACGATCAGCGGCCCTCAGTGGTCCCTGACCGATTCTCTGGCTCCAAGTCCGGCACCTCA GCCTCCCTGGCCATCAGTGGGCTCCAGTCTGAAGATGAGGCCGAATATTACTGTGCAGCATGGGATGACAG CCTGAAGGGGGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-004 VL PROTEIN (SEQ ID NO: 8) QSVLTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVVPDRFSGSKSGTS ASLAISGLQSEDEAEYYCAAWDDSLKGAVFGGGTQLTVL >SC09-005 VH DNA (SEQ ID NO: 9) CAGGTGCAGCTGGTGCAATCTGGGGCTGAGGTCAAGAGGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATTAGTTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGTCTACGCACAGAAATTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTT CTGTGCGAGGCACGGGAACTATTATTACTACTCCGGTATGGACCTCTGGGGCCAAGGGACCACGGTCACC >SC09-005 VH PROTEIN (SEQ ID NO: 10) QVQLVQSGAEVKRPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTVYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDLWGQGTTVT >SC09-005 VL DNA (SEQ ID NO: 11) CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTGGGTCTCCTGGACAGTCGATCACCATCTCCTGCACTGG AACCAGCAGTGACGTCGGTGGTTATAACTATGTCTCCTGGTACCAACAACACCCAGGCAAAGCCCCCAAAC TCCTGATTTTTGATGTCAGTGATCGGCCCTCAGGGGTTTCTGATCGCTTCTCTGGCTCCAAGTCTGCGGAC ACGGCCTCCCTGACCATCTCTGGACTCCAGGCTCAGGACGAGGCTGATTATTACTGCTGCTCATATGCAGG TAGTGCCAAGGGCGTCTTCGGAACTGGGACCAAGGTCACCGTCCTAG >SC09-005 VL PROTEIN (SEQ ID NO: 12) QSALTQPASVSGSPGQSITISCTGTSSDVGGYNYVSWYQQHPGKAPKLLIFDVSDRPSGVSDRFSGSKSAD TASLTISGLQAQDEADYYCCSYAGSAKGVFGTGTKVTVL >SC09-006 VH DNA (SEQ ID NO: 13) GAGGTGCAGCTGGTGGAGTCTGGGGCTGAGGTCAAGAGGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATTAGTTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGTCTACGCACAGAAATTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTT CTGTGCGAGGCACGGGAACTATTATTACTACTCCGGTATGGACCTCTGGGGCCAAGGGACCACGGTCACC >SC09-006 VH PROTEIN (SEQ ID NO: 14) EVQLVESGAEVKRPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTVYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDLWGQGTTVT >SC09-006 VL DNA (SEQ ID NO: 15) TCCTATGTGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGACAGACGGCCAGGATTACCTGTGGGGG AAACAACATTGGAAGTAAAACTGTGCATTGGTACCAGCAGAACTCAGGCCAGGCCCCTGTGCTGGTCGTCT ATGGTGATAGCGACCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGACCACGGCCACC CTGACCATCAGCAGGGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATAGTAGTAGTGA TCATCCCGGTGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-006 VL PROTEIN (SEQ ID NO: 16) SYVLTQPPSVSVAPGQTARITCGGNNIGSKTVHWYQQNSGQAPVLVVYGDSDRPSGIPERFSGSNSGTTAT LTISRVEAGDEADYYCQVWDSSSDHPGAVFGGGTQLTVL >SC09-007 VH DNA (SEQ ID NO: 17) CAGGTGCAGCTGGTGCAATCTGGAGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGAGGGATCAGCCCTATCTTTGGTTCAGCAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTACC GCGGACATATTTTCGAACACAGTGTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTGTATTA CTGTGCGAGACACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-007 VH PROTEIN (SEQ ID NO: 18) QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSAAYAQKFQGRVTIT ADIFSNTVYMELNSLTSEDTAVYYCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-007 VL DNA (SEQ ID NO: 19) TCCTATGTGCTGACTCAGCCACCCTCAGCGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGG AAGCAGCTCCAACATCGGAAGTAATACTGTAAACTGGTACCAGCAGGTCCCCGGAACGGCCCCCAAACTCC TCATCTATGGTGATGATCAGCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCA GCCTCCCTGGCCATCAGTGGGCTCCAGTCTGAGGATGAGGCTGATTATTACTGTGCAACATGGGATGACAG CCTGAATGGTCATGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-007 VL PROTEIN (SEQ ID NO: 20) SYVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQVPGTAPKLLIYGDDQRPSGVPDRFSGSKSGTS ASLAISGLQSEDEADYYCATWDDSLNGHVFGGGTQLTVL >SC09-008 VH DNA (SEQ ID NO: 21) GAGGTCCAGCTGGTGCAGTCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAGAGTCTCCTGTAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTT CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-008 VH PROTEIN (SEQ ID NO: 22) EVQLVQSGAEVKKPGSSVRVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-008 VL DNA (SEQ ID NO: 23) TCCTATGTGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGACAGACGGCCAGGATTACCTGTGGGGG AAACAACATTGGAAGTAAAACTGTGCATTGGTACCAGCAGAACTCAGGCCAGGCCCCTGTGCTGGTCGTCT ATGGTGATAGCGACCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGACCACGGCCACC CTGACCATCAGCAGGGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATAGTAGTAGTGA TCATCCCGGTGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-008 VL PROTEIN (SEQ ID NO: 24) SYVLTQPPSVSVAPGQTARITCGGNNIGSKTVHWYQQNSGQAPVLVVYGDSDRPSGIPERFSGSNSGTTAT LTISRVEAGDEADYYCQVWDSSSDHPGAVFGGGTQLTVL >SC09-009 VH DNA (SEQ ID NO: 25) CAGGTGCAGCTGGTGCAATCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAATTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGGCATCTGAGGACACGGCCGTATATTT CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-009 VH PROTEIN (SEQ ID NO: 26) QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLASEDTAVYFCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-009 VL DNA (SEQ ID NO: 27) GACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGTCG GGCGAGTCAGCATATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAGCCAGGGAAAGGCCCTCAGCTCCTGA TCTATTCTGCATCCCGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTC ACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGCTAACAGTTTCCC CCTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAAC >SC09-009 VL PROTEIN (SEQ ID NO: 28) DIQMTQSPSSVSASVGDRVTITCRASQHISSWLAWYQQKPGKGPQLLIYSASRLQSGVPSRFSGSGSGTDF TLTISSLQPEDFATYYCQQANSFPLTFGPGTKVDIK >SC09-010 VH DNA (SEQ ID NO: 29) GAGGTGCAGCTGGTGGAGTCCGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAATTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCCAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTA CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-010 VH PROTEIN (SEQ ID NO: 30) EVQLVESGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYYCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-010 VL DNA (SEQ ID NO: 31) TCCTATGTGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGACAGACGGCCAGGATTACCTGTGGGGG AAACAACATYGGAAGTAAAACTGTGCATTGGTACCAGCAGAACTCAGGCCAGGCCCCTGTGCTGGTCGTCT TTGTTGATAGCGACCGTCCCTCAGGGATCCATGAGCGATTCTGTGGCTCCAACTCTGGGTCCACGGCCACC CTGACCATCAGCAGCGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATAGTAATAGCGA TCATCCCGGTGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-010 VL PROTEIN (SEQ ID NO: 32) SYVLTQPPSVSVAPGQTARITCGGNNIGSKTVHWYQQNSGQAPVLVVFVDSDRPSGIHERFCGSNSGSTAT LTISSVEAGDEADYYCQVWDSNSDHPGAVFGGGTQLTVL >SC09-011 VH DNA (SEQ ID NO: 33) GAGGTCCAGCTGGTACAGTCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGGCAGGCCCCTGGACAAGGCCTTGACTGGA TGGGAGGGATCAGCCCTATCTTTGGTTCAGCAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTACC GCGGACATATTTTCGAACACAGTGTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTGTATTA CTGTGCGAGACACGGGAATTATTATTACTACTCCGGTACGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-011 VH PROTEIN (SEQ ID NO: 34) EVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSAAYAQKFQGRVTIT ADIFSNTVYMELNSLTSEDTAVYYCARHGNYYYYSGTDVWGQGTTVTVSS >SC09-011 VL DNA (SEQ ID NO: 35) TCCTATGTGCTGACTCAGCCACCCGCAGTGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCGTGTTCTGG AAGTGATTCCAACATCGGGAGAAGAAGTGTAAACTGGTACCAGCAGTTCCCAGGAACGGCCCCCAAACTCC TCATCTATAGTAACGATCAGCGGCCCTCAGTGGTCCCTGACCGATTCTCTGGCTCCAAGTCCGGCACCTCA GCCTCCCTGGCCATCAGTGGGCTCCAGTCTGAAGATGAGGCCGAATATTACTGTGCAGCATGGGATGACAG CCTGAAGGGGGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-011 VL PROTEIN (SEQ ID NO: 36) SYVLTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVVPDRFSGSKSGTS ASLAISGLQSEDEAEYYCAAWDDSLKGAVFGGGTQLTVL >SC09-012 VH DNA (SEQ ID NO: 37) GAGGTCCAGCTGGTACAGTCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAATTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGAGGGATCAGCCCTATTTTTGGTTCAGCAGTCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTACC GCGGACATATTTTCGAACACAGTGTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTGTATTA CTGTGCGAGACACGGGACTTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-012 VH PROTEIN (SEQ ID NO: 38) EVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSAVYAQKFQGRVTIT ADIFSNTVYMELNSLTSEDTAVYYCARHGTYYYYSGMDVWGQGTTVTVSS >SC09-012 VL DNA (SEQ ID NO: 39) CAGTCTGTCGTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCATCTCCTGCACTGG GAGCAGCTCCAACATCGGGGCAGGTTATGATGTACACTGGTACCAGCAGCTTCCAGGGACAGCCCCCAAAC TCCTCATCTATGGTAACAACAATCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACC TCAGCCTCCCTGGCCATCACTGGGCTCCAGGTTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACCA GAACCTGAGTGAGGGGGTCTTCGGCGGAGGGACCAAGCTGACCGTCCTAG >SC09-012 VL PROTEIN (SEQ ID NO: 40) QSVVTQPPSVSGAPGQRVTISCTGSSSNIGAGYDV SASLAITGLQVEDEADYYCQSYDQNLSEGVFGGGTKLTVL >SC09-029 VH DNA (SEQ ID NO: 41) GAGGTGCAGCTGGTGGAGTCCGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTA CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-029 VH PROTEIN (SEQ ID NO: 42) EVQLVESGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYYCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-029 VL DNA (SEQ ID NO: 43) GAAATTGTGATGACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCTGGGGAAAGAGGCACCCTCTCCTGCAG GGCCAGTCAGAGTGTTAGCAGCTACTTAGCCTGGTACCAACAGAAACCTGGCCAGGCTCCCAGGCTCCTCA TCTATGGTGCATCCACCAGGGCCACTGGCATCCCAGACAGGTTCACTGGCAGTGGGTCTGGGACAGACTTC ACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCAGTATGGGAGCTCACC ATTCGCTTTCGGCCCTGGGACCAAGGTGGAGATCAAA >SC09-029 VL PROTEIN (SEQ ID NO: 44) EIVMTQSPGTLSLSPGERGTLSCRASQSVSSYLAWYQQKPGQAPRLLIYGASTRATGIPDRFTGSGSGTDF TLTISRLEPEDFAVYYCQQYGSSPFAFGPGTKVEIK >SC09-030 VH DNA (SEQ ID NO: 45) CAGATGCAGCTGGTGCAGTCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTT CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-030 VH PROTEIN (SEQ ID NO: 46) QMQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-030 VL DNA (SEQ ID NO: 47) TCCTATGTGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGACAGACGGCCAGGATTACCTGTGGGGG AAACAACATTGGAAGTAAAAGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTGGTCGTCT ATGGTGATAGCGACCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGACCACGGCCACC CTGACCATCAGCAGGGTCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATAGTAGTAGTGA TCATCCCGGTGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-030 VL PROTEIN (SEQ ID NO: 48) SYVLTQPPSVSVAPGQTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYGDSDRPSGIPERFSGSNSGTTAT LTISRVEAGDEADYYCQVWDSSSDHPGAVFGGGTQLTVL >SC09-031 VH DNA (SEQ ID NO: 49) CAGGTCCAGCTGGTACAGTCTGGGGCTGAGGTCGAGAGGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGCGGCACCTCCAATAACTATGCCATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGTCTGACATCTGAGGACACGGCCGTCTATTA TTGTGCGAGACACGGGAATTATTATTACAACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-031 VH PROTEIN (SEQ ID NO: 50) QVQLVQSGAEVERPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYYCARHGNYYYNSGMDVWGQGTTVTVSS >SC09-031 VL DNA (SEQ ID NO: 51) CAGTCTGTGTTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCATCTCCTGCACTGG GAGCAGCTCCAACATCGGGGCAGGTTATGATGTACACTGGTACCAGCAGCTTCCAGAAACAGCCCCCAAAC TCCTCATTTATGATAACAACAATCGTCCCTCAGGGGTTTCTGACCGATTCTCTGGCTCCAAGTCTGGCACT TCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACAG CGGCCTGAGTGCTTCGCCTTATGTCTTCGGAGCTGGGACCAAGGTCACCGTCCTAG >SC09-031 VL PROTEIN (SEQ ID NO: 52) QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDV SASLAITGLQAEDEADYYCQSYDSGLSASPYVFGAGTKVTVL >SC09-112 VH DNA (SEQ ID NO: 53) CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAGTTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTA CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-112 VH PROTEIN (SEQ ID NO: 54) QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYYCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-112 VL DNA (SEQ ID NO: 55) CAGTCTGTGTTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCATCTCCTGCACTGG GAGCAGCGCCAACATCGGGGCAGGTTATGATGTCCACTGGTACCAGCAGTTTCCAGGAACAGCCCCCAAAC TCCTCATCTATGGTAACAACAATCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACC TCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACAG CAGCCTGAGTGGTGCGTTATTCGGCGGAGGGACCAAGCTGACCGTCCTAG >SC09-112 VL PROTEIN (SEQ ID NO: 56) QSVLTQPPSVSGAPGQRVTISCTGSSANIGAGYDV SASLAITGLQAEDEADYYCQSYDSSLSGALFGGGTKLTVL >SC09-113 VH DNA (SEQ ID NO: 57) CAGATGCAGCTGGTGCAGTCTGGGGCTGAGGTCAAGAAGGCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGAGTGGA TGGGCGGGATCAGTCCAATCTTTGGTTCGACAGTCTACGCACAGAAATTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCACACACTGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGCATATTT CTGTGCGAGGCACGGAAACTATTATTACTACTCCGGTATGGACCTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-113 VH PROTEIN (SEQ ID NO: 58) QMQLVQSGAEVKKAGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLEWMGGISPIFGSTVYAQKFQGRVTIS ADIFSHTAYMELNSLTSEDTAAYFCARHGNYYYYSGMDLWGQGTTVTVSS >SC09-113 VL DNA (SEQ ID NO: 59) CAGTCTGTGCTGACTCAGCCACCCGCAGTGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCGTGTTCTGG AAGTGATTCCAACATCGGGAGAAGAAGTGTAAACTGGTACCAGCAGTTCCCAGGAACGGCCCCCAAACTCC TCATCTATAGTAACGATCAGCGGCCCTCAGTGGTCCCTGACCGATTCTCTGGCTCCAAGTCCGGCACCTCA GCCTCCCTGGCCATCAGTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGTGCAGCATGGGATGCCAG CCTGAGTGGTCCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-113 VL PROTEIN (SEQ ID NO: 60) QSVLTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVVPDRFSGSKSGTS ASLAISGLQAEDEADYYCAAWDASLSGPVFGGGTQLTVL >SC09-114 VH DNA (SEQ ID NO: 61) CAGGTGCAGCTGGTGCAATCTGGGGCTGAGGTCAAGAAGCCTGGGTCCTCGGTGAAAGTCTCCTGCAAGTC TTCTGGAGGCACCTCCAATAACTATGCTATCAGCTGGGTGCGACAGGCCCCTGGACAAGGCCTTGACTGGA TGGGCGGGATCAGCCCTATCTTTGGTTCGACAGCCTACGCACAGAAATTCCAGGGCAGAGTCACTATTTCC GCGGACATATTTTCGAACACAGCCTACATGGAGCTGAACAGCCTGACATCTGAGGACACGGCCGTATATTT CTGTGCGAGGCACGGGAATTATTATTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCGAGC >SC09-114 VH PROTEIN (SEQ ID NO: 62) QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGSTAYAQKFQGRVTIS ADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDVWGQGTTVTVSS >SC09-114 VL DNA (SEQ ID NO: 63) TCCTATGTGCTGACTCAGCCACCCGCAGTGTCTGGGACCCCCGGGCAGAGGGTCACCATCTCGTGTTCTGG AAGTGATTCCAACATCGGGAGAAGAAGTGTAAACTGGTACCAGCAGTTCCCAGGAACGGCCCCCAAACTCC TCATCTATAGTAACGATCAGCGGCCCTCAGTGGTCCCTGACCGATTCTCTGGCTCCAAGTCCGGCACCTCA GCCTCCCTGGCCATCAGTGGGCTCCAGTCTGAAGATGAGGCCGAATATTACTGTGCAGCATGGGATGACAG CCTGAAGGGGGCTGTGTTCGGAGGAGGCACCCAGCTGACCGTCCTCG >SC09-114 VL PROTEIN (SEQ ID NO: 64) SYVLTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVVPDRFSGSKSGTS ASLAISGLQSEDEAEYYCAAWDDSLKGAVFGGGTQLTVL Vector pIg-C911-HCgamma1 (SEQ ID NO:175) tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 60 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 120 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 180 gcttagggtt aggcgttttg cgctgcttcg ctaggtggtc aatattggcc attagccata 240 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat 300 ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat 360 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat 420 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 480 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 540 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 600 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 660 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 720 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 780 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 840 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 900 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc 960 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc 1020 ctccgcggcc gggaacggtg cattggaagc tggcctggat atcctgactc tcttaggtag 1080 ccttgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt 1140 taaggagatc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag 1200 gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag 1260 ttcaattaca gctcgccacc atgggatgga gctgtatcat cctcttcttg gtactgctgc 1320 tggcccagcc ggccagtgac cttgaccggt gcaccacttt tgatgatgtt caagctccta 1380 attacactca acatacttca tctatgaggg gggtttacta tcctgatgaa atttttagat 1440 cggacactct ttatttaact caggatttat ttcttccatt ttattctaat gttacagggt 1500 ttcatactat taatcatacg tttggcaacc ctgtcatacc ttttaaggat ggtatttatt 1560 ttgctgccac agagaaatca aatgttgtcc gtggttgggt ttttggttct accatgaaca 1620 acaagtcaca gtcggtgatt attattaaca attctactaa tgttgttata cgagcatgta 1680 actttgaatt gtgtgacaac cctttctttg ctgtttctaa acccatgggt acacagacac 1740 atactatgat attcgataat gcatttaatt gcactttcga gtacatatct gatgcctttt 1800 cgcttgatgt ttcagaaaag tcaggtaatt ttaaacactt acgagagttt gtgtttaaaa 1860 ataaagatgg gtttctctat gtttataagg gctatcaacc tatagatgta gttcgtgatc 1920 taccttctgg ttttaacact ttgaaaccta tttttaagtt gcctcttggt attaacatta 1980 caaattttag agccattctt acagcctttt cacctgctca agacatttgg ggcacgtcag 2040 ctgcagccta ttttgttggc tatttaaagc caactacatt tatgctcaag tatgatgaaa 2100 atggtacaat cacagatgct gttgattgtt ctcaaaatcc acttgctgaa ctcaaatgct 2160 ctgttaagag ctttgagatt gacaaaggaa tttaccagac ctctaatttc agggttgttc 2220 cctcaggaga tgttgtgaga ttccctaata ttacaaactt gtgtcctttt ggagaggttt 2280 ttaatgctac taaattccct tctgtctatg catgggagag aaaaaaaatt tctaattgtg 2340 ttgctgatta ctctgtgctc tacaactcaa catttttttc aacctttaag tgctatggcg 2400 tttctgccac taagttgaat gatctttgct tctccaatgt ctatgcagat tcttttgtag 2460 tcaagggaga tgatgtaaga caaatagcgc caggacaaac tggtgttatt gctgattata 2520 attataaatt gccagatgat ttcatgggtt gtgtccttgc ttggaatact aggaacattg 2580 atgctacttc aactggtaat tataattata aatataggta tcttagacat ggcaagctta 2640 ggccctttga gagagacata tctaatgtgc ctttctcccc tgatggcaaa ccttgcaccc 2700 cacctgctct taattgttat tggccattaa atgattatgg tttttacacc actactggca 2760 ttggctacca accttacaga gttgtagtac tttcttttga acttttaaat gcaccggcca 2820 cggtttgtgg accaaaatta tccactgacc ttattaagaa ccagtgtgtc aattttaatt 2880 ttaatggact cactggtact ggtgtgttaa ctccttcttc aaagagattt caaccatttc 2940 aacaatttgg ccgtgatgtt tctgatttca ctgattccgt tcgagatcct aaaacatctg 3000 aaatattaga catttcacct tgctcttttg ggggtgtaag tgtaattaca cctggaacaa 3060 atgcttcatc tgaagttgct gttctatatc aagatgttaa ctgcactgat gtttctacag 3120 caattcatgc agatcaactc acaccagctt ggcgcatata ttctactgga aacaatgtat 3180 tccagactca ggcaggctgt cttataggag ctgagcatgt cgacacttct tatgagtgcg 3240 acattcctat tggagctggc atttgtgcta gttaccatac agtttcttta ttacgtagta 3300 ctagccaaaa atctattgtg gcttatacta tgtctttagg tgctgatagt tcaattgctt 3360 actctaataa caccattgct atacctacta acttttcaat tagcattact acagaagtaa 3420 tgcctgtttc tatggctaaa acctccgtag attgtaatat gtacatctgc ggagattcta 3480 ctgaatgtgc taatttgctt ctccaatatg gtagcttttg cacacaacta aatcgtgcac 3540 tctcaggtat tgctgctgaa caggatcgca acacacgtga agtgttcgct caagtcaaac 3600 aaatgtacaa aaccccaact ttgaaatatt ttggtggttt taatttttca caaatattac 3660 ctgaccctct aaagccaact aagaggtctt ttattgagga cttgctcttt aataaggtga 3720 cactcgctga tgctggcttc atgaagcaat atggcgaatg cctaggtgat attaatgcta 3780 gagatctcat ttgtgcgcag aagttcaatg gacttacagt gttgccacct ctgctcactg 3840 atgatatgat tgctgcctac actgctgctc tagttagtgg tactgccact gctggatgga 3900 catttggtgc tggcgctgct cttcaaatac cttttgctat gcaaatggca tataggttca 3960 atggcattgg agttacccaa aatgttctct atgagaacca aaaacaaatc gccaaccaat 4020 ttaacaaggc gattagtcaa attcaagaat cacttacaac aacatcaact gcattgggca 4080 agctgcaaga cgttgttaac cagaatgctc aagcattaaa cacacttgtt aaacaactta 4140 gctctaattt tggtgcaatt tcaagtgtgc taaatgatat cctttcgcga cttgataaag 4200 tcgaggcgga ggtacaaatt gacaggttaa ttacaggcag acttcaaagc cttcaaacct 4260 atgtaacaca acaactaatc agggctgctg aaatcagggc ttctgctaat cttgctgcta 4320 ctaaaatgtc tgagtgtgtt cttggacaat caaaaagagt tgacttttgt ggaaagggct 4380 accaccttat gtccttccca caagcagccc cgcatggtgt tgtcttccta catgtcacgt 4440 atgtgccatc ccaggagagg aacttcacca cagcgccagc aatttgtcat gaaggcaaag 4500 catacttccc tcgtgaaggt gtttttgtgt ttaatggcac ttcttggttt attacacaga 4560 ggaacttctt ttctccacaa ataattacta cagacaatac atttgtctca ggaaattgtg 4620 atgtcgttat tggcatcatt aacaacacag tttatgatcc tctgcaacct gagcttgact 4680 cattcaaaga agagctggac aagtacttca aaaatcatac atcaccagat gttgattttg 4740 gcgacatttc aggcattaac gcttctgtcg tcaacattca aaaagaaatt gaccgcctca 4800 atgaggtcgc taaaaattta aatgaatcac tcattgacct tcaagaactg ggaaaatatg 4860 agcaatatat taaatggcct ctcgacgaac aaaaactcat ctcagaagag gatctgaatg 4920 ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc tttaaggtgg 4980 tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt atcatcctca 5040 tcatgctttg gcagaagaag ccacgttagg cggccgctcg agtgctagca ccaagggccc 5100 cagcgtgttc cccctggccc ccagcagcaa gagcaccagc ggcggcacag ccgccctggg 5160 ctgcctggtg aaggactact tccccgagcc cgtgaccgtg agctggaaca gcggcgcctt 5220 gaccagcggc gtgcacacct tccccgccgt gctgcagagc agcggcctgt acagcctgag 5280 cagcgtggtg accgtgccca gcagcagcct gggcacccag acctacatct gcaacgtgaa 5340 ccacaagccc agcaacacca aggtggacaa acgcgtggag cccaagagct gcgacaagac 5400 ccacacctgc cccccctgcc ctgcccccga gctgctgggc ggaccctccg tgttcctgtt 5460 cccccccaag cccaaggaca ccctcatgat cagccggacc cccgaggtga cctgcgtggt 5520 ggtggacgtg agccacgagg accccgaggt gaagttcaac tggtacgtgg acggcgtgga 5580 ggtgcacaac gccaagacca agccccggga ggagcagtac aacagcacct accgggtggt 5640 gagcgtgctc accgtgctgc accaggactg gctgaacggc aaggagtaca agtgcaaggt 5700 gagcaacaag gccctgcctg cccccatcga gaagaccatc agcaaggcca agggccagcc 5760 ccgggagccc caggtgtaca ccctgccccc cagccgggag gagatgacca agaaccaggt 5820 gtccctcacc tgtctggtga agggcttcta ccccagcgac atcgccgtgg agtgggagag 5880 caacggccag cccgagaaca actacaagac caccccccct gtgctggaca gcgacggcag 5940 cttcttcctg tacagcaagc tcaccgtgga caagagccgg tggcagcagg gcaacgtgtt 6000 cagctgcagc gtgatgcacg aggccctgca caaccactac acccagaaga gcctgagcct 6060 gagccccggc aagtgataat ctagagggcc cgtttaaacc cgctgatcag cctcgactgt 6120 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 6180 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 6240 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 6300 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 6360 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 6420 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 6480 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 6540 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 6600 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 6660 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 6720 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 6780 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 6840 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 6900 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 6960 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 7020 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 7080 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 7140 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 7200 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 7260 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 7320 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 7380 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 7440 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 7500 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 7560 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 7620 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 7680 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 7740 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 7800 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 7860 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 7920 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 7980 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 8040 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 8100 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 8160 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 8220 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 8280 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 8340 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 8400 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 8460 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 8520 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 8580 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 8640 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 8700 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 8760 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 8820 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 8880 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 8940 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 9000 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 9060 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 9120 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 9180 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 9240 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 9300 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 9360 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 9420 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 9480 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 9540 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 9600 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 9660 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 9720 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 9780 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 9840 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 9900 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 9960 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 10020 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 10080 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 10140 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 10200 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 10260 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 10320 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 10380 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 10440 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 10500 aagtgccacc tgacg 10515 Vector pIg-C909-Ckappa (SEQ ID NO:176) tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 60 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 120 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgtt aattaacatg 180 aagaatctgc ttagggttag gcgttttgcg ctgcttcgct aggtggtcaa tattggccat 240 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata 300 cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat 360 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 420 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 480 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 540 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 600 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 660 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 720 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 780 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 840 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 900 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc 960 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc 1020 gatccagcct ccgcggccgg gaacggtgca ttggaatcga tgactctctt aggtagcctt 1080 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag 1140 gagatcaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac 1200 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca 1260 attacagctc gccaccatgc ggctgcccgc ccagctgctg ggccttctca tgctgtgggt 1320 gcccgcctcg agatctatcg atgcatgcca tggtaccaag cttgccacca tgagcagcag 1380 ctcttggctg ctgctgagcc tggtggccgt gacagccgcc cagagcacca tcgaggagca 1440 ggccaagacc ttcctggaca agttcaacca cgaggccgag gacctgttct accagagcag 1500 cctggccagc tggaactaca acaccaacat caccgaggag aacgtgcaga acatgaacaa 1560 cgccggcgac aagtggagcg ccttcctgaa ggagcagagc acactggccc agatgtaccc 1620 cctgcaggag atccagaacc tgaccgtgaa gctgcagctg caggccctgc agcagaacgg 1680 cagcagcgtg ctgagcgagg acaagagcaa gcggctgaac accatcctga acaccatgtc 1740 caccatctac agcaccggca aagtgtgcaa ccccgacaac ccccaggagt gcctgctgct 1800 ggagcccggc ctgaacgaga tcatggccaa cagcctggac tacaacgagc ggctgtgggc 1860 ctgggagagc tggcggagcg aagtgggcaa gcagctgcgg cccctgtacg aggagtacgt 1920 ggtgctgaag aacgagatgg ccagggccaa ccactacgag gactacggcg actactggag 1980 aggcgactac gaagtgaacg gcgtggacgg ctacgactac agcagaggcc agctgatcga 2040 ggacgtggag cacaccttcg aggagatcaa gcctctgtac gagcacctgc acgcctacgt 2100 gcgggccaag ctgatgaacg cctaccccag ctacatcagc cccatcggct gcctgcccgc 2160 ccacctgctg ggcgacatgt ggggccggtt ctggaccaac ctgtacagcc tgaccgtgcc 2220 cttcggccag aagcccaaca tcgacgtgac cgacgccatg gtggaccagg cctgggacgc 2280 ccagcggatc ttcaaggagg ccgagaagtt cttcgtgagc gtgggcctgc ccaacatgac 2340 ccagggcttt tgggagaaca gcatgctgac cgaccccggc aatgtgcaga aggccgtgtg 2400 ccaccccacc gcctgggacc tgggcaaggg cgacttccgg atcctgatgt gcaccaaagt 2460 gaccatggac gacttcctga ccgcccacca cgagatgggc cacatccagt acgacatggc 2520 ctacgccgcc cagcccttcc tgctgcggaa cggcgccaac gagggctttc acgaggccgt 2580 gggcgagatc atgagcctga gcgccgccac ccccaagcac ctgaagagca tcggcctgct 2640 gagccccgac ttccaggagg acaacgagac cgagatcaac ttcctgctga agcaggccct 2700 gaccatcgtg ggcaccctgc ccttcaccta catgctggag aagtggcggt ggatggtgtt 2760 taagggcgag atccccaagg accagtggat gaagaagtgg tgggagatga agcgggagat 2820 cgtgggcgtg gtggagcccg tgccccacga cgagacctac tgcgaccccg ccagcctgtt 2880 ccacgtgagc aacgactact ccttcatccg gtactacacc cggaccctgt accagttcca 2940 gttccaggag gccctgtgcc aggccgccaa gcacgagggc cccctgcaca agtgcgacat 3000 cagcaacagc accgaggccg gacagaaact gttcaacatg ctgcggctgg gcaagagcga 3060 gccctggacc ctggccctgg agaatgtggt gggcgccaag aacatgaatg tgcgccccct 3120 gctgaactac ttcgagcccc tgttcacctg gctgaaggac cagaacaaga acagcttcgt 3180 gggctggagc accgactgga gcccctacgc cgaccagagc atcaaagtgc ggatcagcct 3240 gaagagcgcc ctgggcgaca aggcctacga gtggaacgac aacgagatgt acctgttccg 3300 gagcagcgtg gcctatgcca tgcggcagta cttcctgaaa gtgaagaacc agatgatcct 3360 gttcggcgag gaggacgtga gagtggccaa cctgaagccc cggatcagct tcaacttctt 3420 cgtgaccgcc cccaagaacg tgagcgacat catcccccgg accgaagtgg agaaggccat 3480 ccggatgagc cggagccgga tcaacgacgc cttccggctg aacgacaact ccctggagtt 3540 cctgggcatc cagcccaccc tgggccctcc caaccagccc cccgtgagca tctggctgat 3600 cgtgtttggc gtggtgatgg gcgtgatcgt ggtgggaatc gtgatcctga tcttcaccgg 3660 catccgggac cggaagaaga agaacaaggc ccggagcggc gagaacccct acgccagcat 3720 cgatatcagc aagggcgaga acaaccccgg cttccagaac accgacgacg tgcagaccag 3780 cttctgataa tctagaacga gctcgaattc gaagcttctg cagacgcgtc gacgtcatat 3840 ggatccgata tcgccgtggc ggccgcaccc agcgtgttca tcttcccccc ctccgacgag 3900 cagctgaaga gcggcaccgc cagcgtggtg tgcctgctga acaacttcta cccccgggag 3960 gccaaggtgc agtggaaggt ggacaacgcc ctgcagagcg gcaacagcca ggagagcgtg 4020 accgagcagg acagcaagga ctccacctac agcctgagca gcaccctcac cctgagcaag 4080 gccgactacg agaagcacaa ggtgtacgcc tgcgaggtga cccaccaggg cctgagcagc 4140 cccgtgacca agagcttcaa ccggggcgag tgttaataga cttaagttta aaccgctgat 4200 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt 4260 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat 4320 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg 4380 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg 4440 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat 4500 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 4560 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 4620 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 4680 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 4740 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 4800 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gccatttcgg 4860 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa 4920 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 4980 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 5040 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 5100 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 5160 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg 5220 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt 5280 cggatctgat cagcacgtga tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt 5340 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc 5400 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc 5460 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat 5520 tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg 5580 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc 5640 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg 5700 cccattcgga ccacaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat 5760 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt 5820 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct 5880 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt 5940 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt 6000 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc 6060 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact 6120 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga 6180 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc 6240 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag 6300 cactcgtccg agggcaaagg aatagcacgt gctacgagat ttcgattcca ccgccgcctt 6360 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 6420 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg 6480 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 6540 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc 6600 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct 6660 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg 6720 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 6780 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 6840 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 6900 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 6960 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 7020 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 7080 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 7140 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 7200 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 7260 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 7320 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 7380 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 7440 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 7500 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 7560 cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 7620 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 7680 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 7740 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 7800 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 7860 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 7920 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 7980 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 8040 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 8100 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 8160 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 8220 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 8280 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 8340 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 8400 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 8460 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 8520 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 8580 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 8640 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 8700 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 8760 aaaagtgcca cctgacg 8777 Vector pIg-C910-Clambda (SEQ ID NO:177) tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 60 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 120 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgtt aattaacatg 180 aagaatctgc ttagggttag gcgttttgcg ctgcttcgct aggtggtcaa tattggccat 240 tagccatatt attcattggt tatatagcat aaatcaatat tggctattgg ccattgcata 300 cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat 360 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 420 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 480 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 540 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 600 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 660 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 720 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 780 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 840 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 900 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc 960 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc 1020 gatccagcct ccgcggccgg gaacggtgca ttggaatcga tgactctctt aggtagcctt 1080 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag 1140 gagatcaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac 1200 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca 1260 attacagctc gccaccatgc ggttctccgc tcagctgctg ggccttctgg tgctgtggat 1320 tcccggcgtc tcgagatcta tcgatgcatg ccatggtacc aagcttgcca ccatgagcag 1380 cagctcttgg ctgctgctga gcctggtggc cgtgacagcc gcccagagca ccatcgagga 1440 gcaggccaag accttcctgg acaagttcaa ccacgaggcc gaggacctgt tctaccagag 1500 cagcctggcc agctggaact acaacaccaa catcaccgag gagaacgtgc agaacatgaa 1560 caacgccggc gacaagtgga gcgccttcct gaaggagcag agcacactgg cccagatgta 1620 ccccctgcag gagatccaga acctgaccgt gaagctgcag ctgcaggccc tgcagcagaa 1680 cggcagcagc gtgctgagcg aggacaagag caagcggctg aacaccatcc tgaacaccat 1740 gtccaccatc tacagcaccg gcaaagtgtg caaccccgac aacccccagg agtgcctgct 1800 gctggagccc ggcctgaacg agatcatggc caacagcctg gactacaacg agcggctgtg 1860 ggcctgggag agctggcgga gcgaagtggg caagcagctg cggcccctgt acgaggagta 1920 cgtggtgctg aagaacgaga tggccagggc caaccactac gaggactacg gcgactactg 1980 gagaggcgac tacgaagtga acggcgtgga cggctacgac tacagcagag gccagctgat 2040 cgaggacgtg gagcacacct tcgaggagat caagcctctg tacgagcacc tgcacgccta 2100 cgtgcgggcc aagctgatga acgcctaccc cagctacatc agccccatcg gctgcctgcc 2160 cgcccacctg ctgggcgaca tgtggggccg gttctggacc aacctgtaca gcctgaccgt 2220 gcccttcggc cagaagccca acatcgacgt gaccgacgcc atggtggacc aggcctggga 2280 cgcccagcgg atcttcaagg aggccgagaa gttcttcgtg agcgtgggcc tgcccaacat 2340 gacccagggc ttttgggaga acagcatgct gaccgacccc ggcaatgtgc agaaggccgt 2400 gtgccacccc accgcctggg acctgggcaa gggcgacttc cggatcctga tgtgcaccaa 2460 agtgaccatg gacgacttcc tgaccgccca ccacgagatg ggccacatcc agtacgacat 2520 ggcctacgcc gcccagccct tcctgctgcg gaacggcgcc aacgagggct ttcacgaggc 2580 cgtgggcgag atcatgagcc tgagcgccgc cacccccaag cacctgaaga gcatcggcct 2640 gctgagcccc gacttccagg aggacaacga gaccgagatc aacttcctgc tgaagcaggc 2700 cctgaccatc gtgggcaccc tgcccttcac ctacatgctg gagaagtggc ggtggatggt 2760 gtttaagggc gagatcccca aggaccagtg gatgaagaag tggtgggaga tgaagcggga 2820 gatcgtgggc gtggtggagc ccgtgcccca cgacgagacc tactgcgacc ccgccagcct 2880 gttccacgtg agcaacgact actccttcat ccggtactac acccggaccc tgtaccagtt 2940 ccagttccag gaggccctgt gccaggccgc caagcacgag ggccccctgc acaagtgcga 3000 catcagcaac agcaccgagg ccggacagaa actgttcaac atgctgcggc tgggcaagag 3060 cgagccctgg accctggccc tggagaatgt ggtgggcgcc aagaacatga atgtgcgccc 3120 cctgctgaac tacttcgagc ccctgttcac ctggctgaag gaccagaaca agaacagctt 3180 cgtgggctgg agcaccgact ggagccccta cgccgaccag agcatcaaag tgcggatcag 3240 cctgaagagc gccctgggcg acaaggccta cgagtggaac gacaacgaga tgtacctgtt 3300 ccggagcagc gtggcctatg ccatgcggca gtacttcctg aaagtgaaga accagatgat 3360 cctgttcggc gaggaggacg tgagagtggc caacctgaag ccccggatca gcttcaactt 3420 cttcgtgacc gcccccaaga acgtgagcga catcatcccc cggaccgaag tggagaaggc 3480 catccggatg agccggagcc ggatcaacga cgccttccgg ctgaacgaca actccctgga 3540 gttcctgggc atccagccca ccctgggccc tcccaaccag ccccccgtga gcatctggct 3600 gatcgtgttt ggcgtggtga tgggcgtgat cgtggtggga atcgtgatcc tgatcttcac 3660 cggcatccgg gaccggaaga agaagaacaa ggcccggagc ggcgagaacc cctacgccag 3720 catcgatatc agcaagggcg agaacaaccc cggcttccag aacaccgacg acgtgcagac 3780 cagcttctga taatctagaa cgagctcgaa ttcgaagctt ctgcagacgc gtcgacgtca 3840 tatggatccg atatcgccgt ggcggccgca ggccagccca aggccgctcc cagcgtgacc 3900 ctgttccccc cctcctccga ggagctgcag gccaacaagg ccaccctggt gtgcctcatc 3960 agcgacttct accctggcgc cgtgaccgtg gcctggaagg ccgacagcag ccccgtgaag 4020 gccggcgtgg agaccaccac ccccagcaag cagagcaaca acaagtacgc cgccagcagc 4080 tacctgagcc tcacccccga gcagtggaag agccaccgga gctacagctg ccaggtgacc 4140 cacgagggca gcaccgtgga gaagaccgtg gcccccaccg agtgcagcta atagacttaa 4200 gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 4260 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 4320 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 4380 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 4440 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 4500 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 4560 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 4620 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 4680 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 4740 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 4800 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 4860 ttttggccat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 4920 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 4980 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 5040 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 5100 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 5160 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 5220 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 5280 ttgtatatcc attttcggat ctgatcagca cgtgatgaaa aagcctgaac tcaccgcgac 5340 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc 5400 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg 5460 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc 5520 ggccgcgctc ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta 5580 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc 5640 cgctgttctg cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca 5700 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga 5760 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac 5820 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc 5880 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg 5940 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt 6000 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt 6060 cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat 6120 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc 6180 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat 6240 cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg 6300 aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac gagatttcga 6360 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 6420 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 6480 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 6540 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 6600 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 6660 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 6720 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 6780 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 6840 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 6900 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 6960 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 7020 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 7080 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 7140 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 7200 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 7260 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 7320 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 7380 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 7440 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 7500 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 7560 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 7620 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 7680 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 7740 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 7800 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 7860 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 7920 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 7980 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 8040 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 8100 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 8160 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 8220 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 8280 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 8340 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 8400 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 8460 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 8520 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 8580 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 8640 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 8700 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 8760 gcgcacattt ccccgaaaag tgccacctga cg 8792 REFERENCES Air MA (1981), Sequence relationships among the hemagglutinin genes of 12 subtypes of influenza A virus. 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De Kruif J et al. (1995), Rapid selection of cell subpopulation-specific human monoclonal antibodies from a synthetic phage antibody library. Proc Natl Acad Sci USA 92:3938.

Ferguson et al., (2003), Nature 422:428-443.

Fouchier AM et al. (2005), Characterization of a novel influenza A virus hemagglutinin subtype (H16) obtained from black-headed gulls. J Virol 79(5):2814-2822.

The World Health Organization Global Influenza Program Surveillance Network (2005), Evolution of H5N1 Avian Influenza Viruses in Asia. Emerg Infect Dis 11:1515-1521.

In this specification where reference has been made to patent specifications, other external documents, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external documents is not to be construed as an admission that such documents, or such sources of information, in any jurisdiction, are prior art, or form part of the common general knowledge in the art.

In the description in this specification reference may be made to subject matter that is not within the scope of the claims of the current application. That subject matter should be readily identifiable by a person skilled in the art and may assist in putting into practice the invention as defined in the claims of this application.