JPH0371666B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a measuring device that automatically measures enzyme immunity, and in particular, substances bound by reaction (Bound) and substances not bound (Free).
This invention relates to an automatic enzyme immunoassay device capable of separating (B-F separation).

Conventionally, enzyme immunoassay is a complicated measurement method, and although the enzyme activity measurement at the final stage can be easily performed automatically, it is difficult to separate B-F at the operational stage where B-F separation is performed. It was difficult to automate the entire process from a place where there was no such thing.

SUMMARY OF THE INVENTION An object of the present invention is to provide an automatic enzyme immunoassay device that automatically performs an analysis process including B-F separation in an enzyme immunoassay.

The present invention is an apparatus for measuring a substance to be measured in a sample by an antigen-antibody reaction using an enzyme as a labeling substance, and a solid phase on which a substance with specific affinity such as an antigen or an antibody is immobilized is supplied to a desired reaction vessel. A phase supply device, a reaction container transport device that continuously transports reaction containers, a sample dispensing device that dispenses a sample into a desired reaction container, and an enzyme labeling reagent or a reagent for enzyme activity measurement into the desired reaction container. A reagent dispensing device that dispenses reagents, a cleaning device that washes the solid phase and reaction container to remove unbound substances while leaving the solid phase in the reaction container, and a photometer that measures enzyme activity after the reaction. The present invention relates to an automatic enzyme immunoassay device equipped with the following.

FIG. 1 is a schematic diagram of the enzyme immunoreaction. 1 is
This is an immobilized solid phase in which antibody (or antigen) 2a is immobilized on a solid phase such as polystyrene balls or glass beads (A). Serum containing the antigen (or antibody) 2b against the antibody (or antigen) 2a or a sample corresponding thereto is added to the immobilized solid phase 1 and reacted (B). Then, antigen (or antibody) 2b
reacts with the antibody (or antigen) 2a on the immobilized solid phase 1 to produce an antigen (antibody)-antibody (antigen) complex immobilized solid phase 3 (C). This composite solidified solid phase 3
Then, an enzyme-labeled antibody (or antigen) 4, which is an enzyme-labeled antibody (or antigen) identical to the antibody (or antigen) 2a immobilized on the immobilized solid phase 1, is added and reacted (D). As a result, an enzyme-labeled antibody (antigen)-antigen (antibody)-antibody (antigen) complex-immobilized solid phase 5, which is a composite of the enzyme-labeled antibody (or antigen) 4 and the complex-immobilized solid phase 3, is generated. (E). Remove excess enzyme-labeled antibody (or antigen) 4,
The enzyme activity on the immobilized solid phase 5 is measured. This allows the content of antigen (or antibody) in the sample to be determined.

FIG. 2 is a partial diagram showing one embodiment of this automatic measuring machine.

U-shaped tube 6 with a large opening 6a and a small opening 6b
First, an immobilized solid phase 1 is prepared which can be freely taken in and taken out from the large opening 6a and which does not enter the small opening 6b. Add the reagent or buffer solution for the immobilized solid phase 1 through the large opening 6a of the U-shaped tube 6 (F). Next, add a certain amount of the sample to be measured using the sample dispenser 7 (G), and
Stirring is performed by blowing air from b to generate the above-mentioned immobilized solid phase 3 (H). Thereafter, the immobilized solid phase 3 and the U-shaped tube 6 are washed by applying a washing liquid supply device 8 to the large opening 6a and a washing liquid suction device 9 to the small opening 6b (I). Next, the above-mentioned enzyme-labeled antibody (or antigen) 4 is applied to the large opening of the U-shaped tube 6 using the dispenser 10.
Add it from a (J), and stir by blowing air through the small mouth 6b to form the above-mentioned immobilized solid phase 5 (K). After this, the cleaning liquid supply device 8 and the cleaning liquid suction device 9
The immobilized solid phase 5 and the U-shaped tube 6 are washed (L). Next, a reagent for measuring the enzyme activity on the immobilized solid phase 5 is added into the U-shaped tube 6 containing the immobilized solid phase 5 using the reagent dispenser 11 (M). Thereafter, air is blown through the small opening 6b for stirring, and after reaction (N), the reaction solution is transferred into the flow cell 13 by the suction tube 12 (O). The absorbance of this reaction solution is measured to determine the enzyme activity on the immobilized solid phase 5. This makes it possible to know the content of antigen (or antibody) 2 in the sample. Further, after the absorbance measurement, the immobilized solid phase suction device 1 provided at the large mouth part 6a is attached to the immobilized solid phase 5 remaining in the U-shaped tube 6.
4, the cleaning liquid is supplied to the small opening 6b by the cleaning liquid supply device 15 into the U-shaped tube 6, and after cleaning, the cleaning liquid is removed by the suction device 14.

FIG. 3 is an overall view showing an embodiment of the automatic measuring machine according to the present invention, in order to explain FIG. 2 in more detail.

FIG. 3 is a top view of a star tank holding a plurality of U-shaped tubes each having a large opening 6a and a small opening 6b. A reaction tube turret 16 for moving the reaction tubes is provided on the upper surface of the thermostatic chamber, and this turret 16 is a ring-shaped disk that rotates at a predetermined pitch in the direction of the arrow. A fixed amount of solid phase is supplied from the immobilized solid phase supply device 17 to the large mouth portion 6a. Next, the turret 16 is rotated and a fixed amount of sample is added by the sample dispenser 7. This sample dispenser 7
The sample is dispensed from the sample cup 19 of the sampler 18, which rotates at a predetermined pitch in the direction of the arrow, into the large mouth portion 6a of the U-shaped tube by the vertical movement of the syringe 20 and the rotation and vertical movement of the probe 21. It can also be configured to dispense an appropriate reagent or buffer at the same time as or before or after dispensing the sample. 22 is an air pump that sends air to the small opening 6b of the U-shaped tube. Air is blown from the small opening 6b to stir the immobilized solid phase and the sample to cause a complete reaction. Next, the unreacted liquid is discharged from the small opening 6b by the liquid discharge pump 23. At the same time, the cleaning pump 24 supplies a cleaning liquid from the large opening 6a to clean the solid phase and the U-shaped tube. The cleaning liquid is discharged from the liquid discharge pump 2.
Discharge according to 3. A vertically movable disc (not shown) is provided on the upper surface of the mouth part 6b, and when air and liquid are supplied and discharged through the mouth part 6b, this disc descends and the supply/discharge tube is connected to the mouth part 6b. The turret 16 is arranged so that it rises when it rotates.

Next, the enzyme-labeled antibody is dispensed from the large mouth portion 6a using the dispenser 10. In the same manner as described above, air is blown into the mouth portion 6b from the air pump 22 for stirring, and the liquid is discharged by the liquid discharge pump 23. The cleaning pump 24 supplies the cleaning liquid from the large opening 6a, and after cleaning the solid phase and the U-shaped tube, the cleaning liquid is sent to the liquid discharge pump 23.
It is discharged by

Next, a reagent is added from the large opening 6a using the reagent dispenser 11, and air is blown into the small opening 6b from the air pump 22 for stirring. Next, the reaction solution is transferred from the small opening 6b to the flow cell 26 by the vacuum pump 25.
Transfer to. Absorbance is measured using this flow cell 26. 27 is a light source, 28 is a filter, and 29 is a light receiving element.

After the measurement, the contents of the U-shaped tube are discharged by a suction device 14 that can be tightly connected to the large opening 6a, and the contents of the U-shaped tube are removed from the small opening 6b.
After cleaning the U-shaped tube by supplying the cleaning liquid by the cleaning liquid supply device 15 attached to the holder, the cleaning liquid is removed by the suction device 14.

Although the enzyme immunoreaction was carried out using a U-shaped tube in the above example, the U-shaped tube is not limited to the U-shaped tube. It is possible to use a shape such as a funnel shape, a pipe having a mesh-like partition inside the pipe, etc.

As described above, according to the present invention, all reactions necessary for sample analysis, removal of substances unbound to the solid phase, etc. can be carried out without changing the reaction vessel while the solid phase is accommodated in the reaction vessel. It is now possible to perform a series of operations, reducing the number of reaction vessels required for analysis, thereby reducing the size of automatic enzyme immunoassay machines and eliminating the complicated transfer of solid phases from one vessel to another. Operation can be omitted. Furthermore, in the present invention, the supply of the solid phase into the reaction vessel, the separation of reactants and unreacted substances, and the measurement of enzyme activity after the reaction can be automatically operated.

[Brief explanation of drawings]

FIG. 1 is a schematic diagram of the enzyme immunoreaction, FIG. 2 is a diagram showing the sequential steps in the automatic measuring device according to the present invention, and FIG. 3 shows the configuration of the automatic measuring device shown in FIG. 2 in more detail. It is a diagram. 1...immobilized solid phase, 2...antigen (antibody), 3...
...Antigen-antibody complex immobilized solid phase, 4...Enzyme labeled antibody, 5...Enzyme labeled antibody-antigen-antibody complex immobilized solid phase, 6...U-shaped tube, 7...Sample pipettor, 8 …
...Cleansing water supply device, 9...Cleansing water suction device, 10
...dispenser, 11 ... reagent dispenser, 12 ... suction tube, 13 ... flow cell, 14 ... immobilized solid phase suction device, 15 ... small mouth washing water supply device, 16 ... reaction Pipe turret, 22...air pump, 23...liquid discharge pump, 24...cleaning pump.

Claims (1)

[Claims] 1. A solid phase supply device that supplies a solid phase immobilized with a specific affinity substance such as an antigen or an antibody to a desired reaction container in a device that measures a substance to be measured in a sample through an antigen-antibody reaction using an enzyme as a labeling substance. , a reaction container transport device that continuously transports reaction containers, a sample dispensing device that dispenses a sample into a desired reaction container, and a reagent for enzyme labeling reagent or enzyme activity measurement reagent into the desired reaction container. Equipped with a reagent dispensing device for dispensing, a cleaning device for washing the solid phase and reaction vessel to remove unbound substances while leaving the solid phase in the reaction vessel, and a photometry device for measuring enzyme activity after the reaction. An automatic enzyme immunoassay device characterized by: