JPS61271455A - Immunological analysis - Google Patents

Abstract

PURPOSE:To enable analysis of an incomplete antibody with an agglutination dispensing with any cleaning process while miniaturizing the equipment, by bonding complements together after activated by being acted on an immunological complex to perform an agglutination of blood cells. CONSTITUTION:A blood cell 2 having an antigen 1 which is allowed to react specifically with an incomplete antibody to be analyzed, complements (C1) 3 which will be activated by being bonded to an immunological complex through an antigen-antibody reaction between an antigen 1 and the incomplete antibody and an anti C1 antibody 4 which is used to bond the complements to be activated by being reacted therewith are arranged as reagents and these reagents are made to react with a serum sample. Then, when any incomplete antibody 5 to be analyzed exists in the sample, it bonds to the antigen 1 by an antigen- antibody reaction and the complements 3 are coupled to the resulting antigen- antibody complex to be activated and the complements thus activated get together through the anti C1 antibody 4 to agglutinate the blood cells 2. Thus, after the reaction between the reagents and the sample, the agglutination and non-agglutination are measured to identify the incomplete antibody 5.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an immunological analysis method, and particularly to an immunological analysis method in which a test substance is analyzed by an agglutination reaction using complement.

[Conventional technology]

Although incomplete antibodies bind to antigens, they do not have the ability to aggregate due to their antigen binding valence, size, or electrical properties of the antigen surface, and therefore cannot be analyzed by ordinary agglutination reactions. Therefore, the Coombs test, which is known as an antiglobulin test, has been used to analyze such incomplete antibodies.

The Coombs test is a red blood cell agglutination test in which red blood cells coated with an incomplete antibody are agglutinated by adding a foreign antibody against that antibody, and is often used as a technique for antibody screening.

[Problem that the invention seeks to solve]

However, in the Coombs test, regardless of the direct or indirect method, antiglobulin serum cannot be used unless red blood cells sensitized with incomplete antibodies are washed several times with a washing solution (physiological saline) to create a blood cell suspension. (For example, anti-1gG antibody), this reacts with immunoglobulin in the fluid other than the sensitized red blood cells, and the desired incomplete antibody (IgG
) cannot be analyzed with high precision. For this reason,
This Coombs test requires a large amount of washing solution to thoroughly wash the red blood cells and create a suspension of the red blood cells.Therefore, when automating this test, a washing device is required and the device becomes large. be.

This invention was made by focusing on these conventional problems, and it is an immunoassay that allows incomplete antibodies to be analyzed with high precision by agglutination reaction without a washing process, and that allows the device to be made compact. The purpose of this website is to provide the following methods:

[Means and Effects for Solving the Problems J] In order to achieve the above object, the present invention uses blood cells that have been sensitized with the incomplete antibodies to be analyzed to complement the immune complexes to which the incomplete antibodies are bound. The complement is activated by the action of the body, and the activated complements bind to each other, causing a blood cell agglutination reaction and analyzing incomplete antibodies.

〔Example〕

The drawing is a schematic reaction diagram showing an example of the present invention. In this example, it is analyzed whether a predetermined incomplete antibody is produced in serum. For this reason, antigen 1 (
Complement (C1) 3 (e.g., fresh guinea pig serum) binds to and activates the immune complex resulting from the antigen-antibody reaction between antigen 1 and incomplete antibodies, and the activated Anti-C1 antibody 4, which binds activated complements in response to complement 3, is used as a reagent, and these reagents and sample serum are reacted, for example, in a reaction container with an inclined bottom surface. .

Here, if there are incomplete antibodies 5 to be analyzed in the sample, these incomplete antibodies 5 bind to antigen 1 through an antigen-antibody reaction, and complement 3 binds to the antigen-antibody complex and is activated. , the activated complements become anti-C1 antibody 4.
The blood cells 2 are bound to each other and agglomerate.

On the other hand, if there is no incomplete antibody 5 to be analyzed in the sample, an antigen-antibody complex for binding and activation by complement 3 will not be formed, so blood cells 2 will not aggregate.

After the reagent and sample are reacted in this manner, agglutination and non-aggregation of the reaction solution are measured by a known method, thereby identifying incomplete antibody 5.

In the above reaction, various reagents and samples may be injected into the reaction container at the same time, or the anti-C1 antibody 4 may be injected after the blood cells 2 having antigen 1 and complement 3 are reacted with the sample. etc., can be injected at any timing.

According to this example, blood cells 2 having antigen 1, blood cells 2 having antigen 1,
An agglutination reaction can be performed simply by injecting complement 3, anti-C1 antibody 4, and a sample into a reaction container. Therefore, a decrease in accuracy due to insufficient washing of blood cells does not become a problem, and a desired incomplete antibody can be analyzed with high accuracy. Furthermore, when configuring an apparatus for carrying out this analysis method, a blood cell washing apparatus is not required, so that the apparatus can be made simple and compact.

Note that the present invention is not limited to the above-described embodiments, and can be modified in many ways. For example, complement 3 does not necessarily need to be separately injected as a reagent, and it is possible to use complement 3 contained in the serum sample. Furthermore, in the above-mentioned embodiment, a so-called indirect method for analyzing whether or not a predetermined incomplete antibody is produced in serum has been described. It can also be effectively applied to the so-called direct method, which analyzes whether the In this case, complement 3 and anti-C1 antibody 4 may be used as reagents, and these reagents may be reacted with blood cells as a sample. Furthermore, the present invention can be effectively applied not only to analyzing the presence or absence of a predetermined incomplete antibody but also to antibody screening and Rh blood type determination.

〔Effect of the invention〕

As described above, according to the present invention, incomplete antibodies can be analyzed by agglutination reaction without washing blood cells as in the Coombs test, which allows for highly accurate analysis and simple equipment. And it can be made small.

[Brief explanation of drawings]

The drawing is a schematic reaction diagram showing an example of the present invention. 1... Antigen 2... Blood cell 3... Complement
4... Anti-C1 antibody 5... Incomplete antibody

Claims (1)

[Claims] 1. Complement is applied to blood cells to which the incomplete antibody to be analyzed is bound, or to immune complexes to which the incomplete antibody to be analyzed is bound. 1. An immunological analysis method comprising: activating a blood cell, binding the activated complements to agglutinate the blood cells, and analyzing the incomplete antibody by measuring the agglutination.