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Assay methods using anti-immune complex antibodies

An immunoassay and complex technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, biological testing, etc., can solve the problems of low reaction efficiency, decreased reaction efficiency, low affinity, etc., and achieves inhibition of cross-reaction. , the effect of slowing down the decline in reaction efficiency and reducing the impact

Pending Publication Date: 2022-07-12
TOSOH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reaction between an antibody immobilized on a solid phase and an antigen proceeds in a heterogeneous system such as a solid-liquid interface, so the reaction efficiency is low (Non-Patent Document 2)
When using a low-affinity mouse monoclonal antibody as a solid phase, it is considered that the reaction at the solid-liquid interface becomes the rate-limiting step and reduces the overall reaction efficiency.

Method used

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  • Assay methods using anti-immune complex antibodies
  • Assay methods using anti-immune complex antibodies
  • Assay methods using anti-immune complex antibodies

Examples

Experimental program
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Effect test

Embodiment 1

[0080] Hereinafter, the method for carrying out the examples of the present invention using rabbit monoclonal antibodies against estradiol (E2) and anti-immune complex mouse monoclonal antibodies against their immune complexes will be described in detail, but the present invention is not limited by these Examples are limited.

[0081] (1) Anti-E2 rabbit monoclonal antibody

[0082] The anti-E2 rabbit monoclonal antibody was obtained by the method described in Japanese Patent Laid-Open No. 2009-240300.

[0083] (2) Anti-immune complex mouse monoclonal antibody

[0084] Antibodies against the immune complex of E2 and anti-E2 rabbit monoclonal antibody (anti-immune complex mouse monoclonal antibody) were obtained by the method described in Japanese Patent No. 6031944 .

[0085] (3) Measurement

[0086] The automatic analysis was performed according to the following method using an automatic chemiluminescence enzyme immunoassay apparatus (AIA-CL2400, manufactured by Tosoh Corpo...

Embodiment 2

[0097] Hereinafter, the method for carrying out the examples of the present invention using a rabbit monoclonal antibody against thyroxine (FT4) and an anti-immune complex mouse monoclonal antibody against their immune complex will be described in detail, but the present invention is not limited to these examples. .

[0098] (1) Anti-FT4 rabbit monoclonal antibody

[0099] The anti-FT4 rabbit monoclonal antibody was obtained by the method described in Japanese Patent Laid-Open No. 2009-240300.

[0100] (2) Anti-immune complex mouse monoclonal antibody

[0101] An antibody against an immune complex of FT4 and an anti-FT4 rabbit monoclonal antibody (anti-immune complex mouse monoclonal antibody) was obtained by the method described in Japanese Patent No. 6031944 .

[0102] (3) Measurement

[0103] The automatic analysis was performed according to the following method using an automatic chemiluminescence enzyme immunoassay apparatus (AIA-CL2400, manufactured by Tosoh Corporati...

Embodiment 3

[0113] Hereinafter, the method for implementing the examples of the present invention using a rabbit monoclonal antibody against estradiol (E2) and an anti-immune complex mouse monoclonal antibody against their immune complexes and using a 96-well ELISA plate will be described in detail, However, the present invention is not limited to these examples.

[0114] (1) Anti-E2 rabbit monoclonal antibody

[0115] As the anti-E2 rabbit monoclonal antibody, the same antibody as described in Example 1 was used.

[0116] (2) Anti-immune complex mouse monoclonal antibody

[0117] As the anti-immune complex mouse monoclonal antibody, the same antibody as described in Example 1 was used.

[0118] (3) Measurement

[0119] The measurement of E2 was performed by the following method using a microplate reader (TECAN, infinite F500).

[0120] Anti-E2 rabbit monoclonal antibody was immobilized at 0.25 μg / mL (carbonate buffer (0.05M pH 9.6)) in a 96-well microplate (black, Greiner). Then, bl...

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Abstract

Provided is a measurement method with which it is possible to improve reaction efficiency in a measurement system using an anti-immune complex antibody. An immunoassay method using an anti-hapten rabbit monoclonal antibody immobilized on a water-insoluble carrier, and a labeled anti-immune complex antibody, the immunoassay method comprises the following steps (i) to iii: (i) a step for reacting an anti-hapten rabbit monoclonal antibody immobilized in a water-insoluble carrier with a hapten in a solution to be assayed; (ii) a step for reacting the labeled anti-immune complex antibody with an immune complex of a hapten-anti-hapten rabbit monoclonal antibody; and (iii) a step for detecting a signal from the marker.

Description

technical field [0001] The present invention relates to assays using anti-immune complex antibodies. Background technique [0002] A hapten, which is a low molecular weight compound, is usually measured by an immunoassay method called a competition method. The competition method is a method in which a hapten contained in a sample and a labeled hapten labeled with a radioisotope, an enzyme, or the like are subjected to a competitive reaction with a certain amount of anti-hapten antibody. When the amount of hapten contained in the sample increases, the amount of binding between the labeled hapten and the anti-hapten antibody decreases. Thereby, the amount of hapten contained in the sample can be estimated based on the binding ratio of the labeled hapten to the anti-hapten antibody. The sensitivity of the assay in the competition method depends on the affinity constant of the anti-hapten antibody used. However, since it is difficult to obtain an anti-hapten antibody with a h...

Claims

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Application Information

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IPC IPC(8): G01N33/53C12Q1/42G01N33/543
CPCG01N33/74G01N2470/06G01N33/54313G01N33/543G01N33/577
Inventor 武藤悠三泽孝一
Owner TOSOH CORP
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