JPS6161059B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an automatic analyzer that automatically measures various proteins efficiently and with high precision by immunoelectrophoresis using agar gel.

Immunoelectrophoresis, which is commonly used for protein analysis, has the advantage of being able to simultaneously analyze many types of proteins and also to identify specific proteins at the same time. However, there were drawbacks such as the need for skill in the preparation of an agarose gel-like support for electrophoresis and immunodiffusion, and in the preparation of holes for sample injection into this support and grooves for injection of antiserum. That is, in the conventional immunoelectrophoresis method, holes for sample injection and grooves for antiserum injection are provided in a gel-like plate of agarose or agar (agar) as a support for electrophoresis and diffusion. It is tightly fixed on a glass plate or plastic film base. This support is placed in a bridging manner between two electrically insulated electrophoresis tanks containing a buffer solution via a connecting material such as a sponge impregnated with the buffer solution, and the support is energized. Performing electrophoresis. The connecting material is not limited to sponge.
A method has also been used in which one end of a piece of filter paper or agarose is brought into contact with a support and the other end is immersed in a buffer solution for electrical connection. The specimen is injected into the specimen injection hole in advance, and the specimen injection hole and the antiserum injection groove are prepared by the analyst or the like before operation. After electrophoresis is complete, when antiserum is injected into the antiserum injection groove, a precipitation line appears due to the reaction between the antigen in the sample and the antibody in the antiserum, and this precipitation line can be optically traced to determine the type and content of the protein. quantity is determined. Desalination, staining, photography, etc. are also performed as necessary. The conventional immunoelectrophoresis described above has various drawbacks as described below.

(1) Skill is required to create the support (dissolve powder such as agarose, pour it onto a glass plate or plastic film, and harden it into a gel-like state) and create holes for sample injection and grooves for antiserum injection, so that uniformity is always achieved. It is difficult and time-consuming to create a suitable support.

(2) Migration unevenness tends to occur due to poor contact when electricity is applied to the support.

(3) It is difficult to adopt a cooling method to prevent heat generation and drying during electrophoresis.

(4) The support easily peels off from the glass plate or plastic film surface and penetrates into the gap through the sample injection hole or antiserum injection groove, which may prevent accurate electrophoresis and immunodiffusion.

The present invention was developed in view of the above points, and is a method for continuously and automatically processing a support having a gel layer of agar, agarose, etc. formed on a transparent film base and provided with a thin removable film cover. drilling, groove cutting, sample dispensing, electrophoresis, antiserum dispensing,
By configuring the process such as immunodiffusion and optical reading to be carried out continuously and automatically, the above-mentioned drawbacks have been overcome; in other words, a uniform support can always be created, and it is possible to The objective is to provide an automatic analyzer that can prevent uneven electrophoresis due to poor contact during electrophoresis, prevent drying of the support, and thus perform accurate electrophoretic diffusion. be.

Hereinafter, the configuration of the present invention will be explained based on embodiments shown in the drawings. FIG. 1 shows an example of a support used in the present invention. This support 1 is constructed by forming a gel layer 3 of agar, agarose, etc. on a transparent film base 2, and covering the top surface of the gel layer 3 with a removably thin film cover 4. The thickness of the gel layer 3 is usually around 1 to 5 mm. The width of the support 1 is usually 7 to 10 cm, and as shown in FIG. 2, the support 1 is stored in a storage box 5 in a rolled state.

FIG. 3 shows an embodiment of an automatic analyzer according to the present invention. A punching device 7 for punching a sample injection hole 6 in a support 1 pulled out from a box by a drive roller described later; a sample dispensing device 8 for dispensing a sample into the sample injection hole 6; Support 1
a groove cutting device 11 for drilling an antiserum injection groove 10 in the sample injection hole 6; and a groove cutting device 11 for drilling a groove 10 for antiserum injection into the sample injection hole 6, and a groove cutting device 11 for performing electrophoresis while bending the support body into which a sample has been dispensed into the sample injection hole 6 into an arc shape by a guide roller to be described later. An electrophoresis tank 12, an antiserum dispensing device 13 for injecting antiserum into the antiserum injection groove 10 of the support after electrophoresis, and a thermostat for performing immunodiffusion of the support into which the antiserum has been dispensed. Humidity tank 1
4, an optical reading device 15 for reading sedimentation lines after diffusion, and a plurality of drive rollers 16 for transporting the support between the above devices.

In the migration tank 12, the support 1 is supported by guide rollers so that its cross section is curved in an arc, as shown in FIG. That is, both ends of the support 1 are immersed in the buffer solution 22 in the electrically insulated buffer solution tanks 17 and 18, respectively.
Both ends of the support 1 are held between guide rollers 20, and the center portion of the lower surface of the support 1 is pushed up by another guide roller 21, so that the support 1 can pass through the migration tank 12 in an arcuate state. It consists of Therefore, as described above, both ends of the support 1 are in direct contact with the buffer solution 22, and the voltage from the electrodes 23 is applied to the support 1 via the buffer solution 22.

In the automatic analyzer of the present invention configured as described above, the support 1 pulled out from the storage box 5
The hole 6 for sample injection is punched together with the film cover 4 by the punching device 7, and then the groove cutting device 1
1, an antiserum injection groove 10 is bored. The cut film cover 4 and gel layer 3 are
It is configured to be sucked into the punching device 7 and the groove cutting device 11 at the same time as the hole or groove is bored. Sample injection hole 6 and antiserum injection groove 10
is formed to a depth that reaches the top surface of the film base 2. Next, after the sample is dispensed into the sample injection hole 6 by the sample dispensing device 8, the support is transferred to the electrophoresis tank 12.
be introduced within. In the migration tank 12, the support 1 is bent by guide rollers 20 and 21 so that its cross section becomes an arc, as shown in FIG. immersed in a buffer solution. When a voltage is applied through the electrode 23, the voltage is applied to the support 1 through the buffer solution 22, and migration is performed. Next, after the antiserum is uniformly injected into the antiserum injection groove 10 by the antiserum dispensing device 13, a reaction by immunodiffusion takes place in a constant temperature and humidity bath 14 whose temperature and humidity are adjusted. That is, a sedimentation line is generated within the gel layer due to the immunoreaction between the antigen in the specimen and the antibody in the antiserum. This sedimentation line is read by an optical reader 15 provided adjacent to and after the constant temperature humid bath 14.
Note that when tracking the movement of the sedimentation line and tracking and reading the concentration caused by tracing the sedimentation line, use the constant temperature humidity tank 14.
An optical reading device may be movably installed within the chamber, or a plurality of constant temperature and humidity baths and a plurality of optical reading devices may be provided. In addition, since the film cover 4 on the surface of the support can be peeled off, it can be washed with water after measurement.
It is possible to dye, photograph, dry, etc. as necessary, and it is also possible to cut and store. Furthermore, since the support 1 is covered with the film cover 4, the water in the gel layer 3 is prevented from evaporating, but by storing it in the storage box 5, evaporation is further prevented. This prevents the gel layer from peeling off.

In the embodiment described above, the groove cutting device 11 is installed in front of the migration tank 12, but as shown in FIG. The apparatus 11 may be installed so that the antiserum injection groove is drilled after electrophoresis is completed.

Since the present invention is configured as described above, there is no need for pre-processing such as creating a support, a hole for specimen injection, and a groove for antiserum injection, and there is no room for individual differences among workers. Further, since the support is energized while being in direct contact with the buffer solution, there is an effect that uneven migration does not occur. Protein analysis by immunoelectrophoresis is a relatively long operation, but by automating it, combined with the above effects, efficient and highly accurate analysis becomes possible.

[Brief explanation of the drawing]

FIG. 1 is a cross-sectional explanatory diagram showing an example of a support used in the automatic analyzer of the present invention, FIG. 2 is a perspective view showing an example of a storage box for the support, and FIG. FIG. 4 is a cross-sectional explanatory diagram showing one embodiment of the present invention, FIG. 4 is a cross-sectional explanatory diagram showing another embodiment of the automatic analyzer of the present invention, and FIG. 5 is a cross-sectional explanatory diagram showing an example of an electrophoresis tank. 1...Support, 2...Film base, 3...
gel layer, 4... film cover, 5... storage box,
6... Sample injection hole, 7... Punching device, 8... Sample dispensing device, 10... Antiserum injection groove, 11...
Grooving device, 12...Migration tank, 13...Antiserum dispensing device, 14...Thermostatic humid tank, 15...Optical reader, 16...Drive roller, 17, 18...Buffer tank, 20 , 21... Guide roller, 22... Buffer solution, 23... Electrode.

Claims (1)

[Claims] 1. A support formed by forming a gel layer of agar, agarose, etc. on a transparent film base and covering the top surface of this gel layer with a removable film cover, and a storage box for storing this support in a rolled state. , a punching device for punching a sample injection hole in a support pulled out from the storage box; a sample dispensing device for dispensing the sample into the sample injection hole;
A groove cutting device for making a groove for antiserum injection in a support, and an electrophoresis tank for performing electrophoresis while bending the support into an arc shape with a guide roller after dispensing the sample into the sample injection hole. , an antiserum dispensing device for injecting antiserum into the antiserum injection groove of the support after electrophoresis, a constant temperature humid bath for performing immunodiffusion of the support into which the antiserum has been dispensed, and a 1. An automatic analyzer comprising: an optical reader for reading sedimentation lines, etc.; and a plurality of drive rollers for transporting a support.