Sandwith immunoassay method and method for detecting antigen by using the same

A technology of sandwich immunoassay and detection method, which is applied in the field of sandwich immunoassay, which can solve the problems of low antigen affinity, difficult high-sensitivity detection, and few antigens

Inactive Publication Date: 2009-05-20
SYSMEX CORP
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] However, the traditional sandwich method has the problem that it is difficult to perform high-sensitivity measurement when the affinity of the antibody used for the measurement to the antigen is not too high and when there is very little antigen in the specimen

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  • Sandwith immunoassay method and method for detecting antigen by using the same

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Embodiment 1

[0103] Using the sandwich immunoassay method of the present invention, the thyroid-stimulating hormone (TSH) is measured according to the following steps.

[0104] Antibodies were diluted with antibody buffer (0.1M MES (2-nitroxanesulfonic acid) (pH 6.0), 1% bovine serum albumin, 0.15M NaCl, 0.1% sodium azide) in the following order.

[0105] 1) Add 30 μL of alkaline phosphatase (ALP)-labeled anti-TSH monoclonal antibody (0.4 U / mL, manufactured by Medix Biochemica) in the reaction vessel as the primary antibody labeled with the marker, and add TSH antigen solution (concentration 0 or 2 μIU / mL, manufactured by Fitzgerald Co.) 30 μL was used as a sample, and reacted at 42° C. for 2 minutes.

[0106] 2) Add 30 μL of streptavidin magnetic particles (average particle size 2 μm, 10 mg / mL in 20 mM sodium phosphate buffer (pH 7.5)), and react at 42° C. for 2.5 minutes.

[0107] 3) Add 30 μL of the biotin-labeled anti-TSH monoclonal antibody (10 μg / mL) as a secondary antibody capable...

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Abstract

The invention provides a sandwich immunoassay method comprising steps of, (a) forming a complex of an antigen and a first antibody by contacting the antigen with the first antibody which recognizes the antigen and is labelled by a detectable labelling substance; and (b) fixing the complex formed in the step (a) to a solid phase by using a second antibody which recognizes the antigen and is capable of binding to the solid phase, as well as a method for detecting an antigen in an analyte by using a method of sandwich immunoassay.

Description

Technical field: [0001] The invention relates to a sandwich immunoassay method capable of sensitively detecting a small amount of antigen contained in a specimen sample and a method for detecting the antigen by the method. Background technique: [0002] A sandwich method is known as a method for detecting a substance (antigen) contained in a sample. The sandwich method is to insert an antigen between two antibodies that can recognize different epitopes of the antigen to detect the antigen. Generally, the sandwich method first allows the antigen to bind to the primary antibody that has been bound to the solid phase. In this way, a complex of the antigen and the primary antibody is formed on the solid phase. The antigen on the solid phase is then detected by allowing the secondary antibody labeled with a detectable label to bind to the antigen. [0003] For example, Patent Publication No. Showa 57-074663 discloses a method for the determination of TSH. This method uses an e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74G01N33/53
CPCG01N33/76G01N33/54306
Inventor 藤本裕之松江美幸
Owner SYSMEX CORP
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