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Chemiluminescence immunoassay kit for detecting anti-SS-B antibody by double-antigen sandwich method

A chemiluminescence immunization, double antigen sandwich technology, applied in the field of medical immunization, can solve problems such as non-specific reactions, and achieve the effect of good repeatability and improved sensitivity

Pending Publication Date: 2021-12-07
捷和泰(北京)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The disadvantage of the indirect method is that the sample needs to be diluted and detected, and the secondary antibody conjugated with the chemiluminescence label will have a non-specific reaction

Method used

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  • Chemiluminescence immunoassay kit for detecting anti-SS-B antibody by double-antigen sandwich method
  • Chemiluminescence immunoassay kit for detecting anti-SS-B antibody by double-antigen sandwich method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of SS-B antigen-coated magnetic beads

[0019] Take 1ug of carboxylated magnetic particles (particle size 1um-3um) suspension, magnetically separate to remove the supernatant, resuspend with 200ul0.1MpH5.0-7.0MES buffer, add 5-10ul freshly prepared 10mg / ml EDC aqueous solution , suspend at room temperature for 30 minutes, add 200ul 0.1M pH5.0-7.0 MES buffer to resuspend after magnetic separation, add 2-10ugSS-B antigen 1, suspend at room temperature for 3-5h, remove supernatant by magnetic separation, and use 2 The solution of %BSA was resuspended to 200ul, suspended at room temperature for 2h, and then resuspended to 200ul with 0.1M pH7.2-8.0 Tris buffer containing 2%BSA to obtain magnetic particles coated with SS-B antigen 1, and stored at 4°C.

Embodiment 2

[0020] The preparation method of embodiment 2 sample diluent

[0021] The buffer system is 0.02M pH7.2 Tris-Hcl, containing 0.03% NaCl, and 0.5% bovine serum albumin.

Embodiment 3

[0022] The preparation method of embodiment 3 SS-B antigen 1 and acridinium ester conjugate

[0023] Take 50ug SS-B antigen, add 250ul 0.1M pH7.0-8.0PBS, mix well, add 1-10ul 5mM acridinium ester, suspend for 30min at room temperature in the dark, add 100ul 1% lysine solution, keep in room temperature in the dark Suspend for 30min. Use a centrifugal desalting column for desalting treatment, and finally collect the liquid in the centrifuge tube into a centrifuge tube, and add an equal volume of glycerol to obtain 800ul SS-B antigen-labeled acridinium ester, and store it in the dark at -20°C.

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Abstract

The invention discloses a chemiluminescence immunodetection kit for detecting an anti-SS-B antibody by a double-antigen sandwich method. An anti-SS-B antibody quality control product and a chemiluminescence substrate solution are included. The kit is characterized by further comprising SS-B antigen coated magnetic beads and an SS-B antigen labeled chemiluminescence marker. The double-antigen sandwich method is used, non-specific reaction in the reaction process is avoided, the two-step reaction is specific reaction, unnecessary non-specific reaction is avoided, the problem of a reaction platform existing in the sandwich method is effectively solved, and the sensitivity can be higher than that of an indirect method. In addition, by increasing the sample adding amount, sample dilution is not needed, and it is guaranteed that CV is better.

Description

technical field [0001] The invention relates to a chemiluminescence immunoassay kit for detecting anti-SS-B antibody by a double-antigen sandwich method, which belongs to the field of medical immunization. Background technique [0002] Chemiluminescent reagents The main detection methodology for self-immunity projects is the indirect method (magnetic beads coupled to antigen, chemiluminescent markers coupled to anti-human IgG). [0003] The advantage of the indirect method is that it is universal. For the self-immunity project of detecting antibodies, only a specific antigen needs to be coupled to the magnetic beads, and the chemiluminescent label is coupled to the secondary antibody. [0004] The disadvantage of the indirect method is that the sample needs to be diluted and detected, and the secondary antibody conjugated with the chemiluminescent label will have a non-specific reaction. Contents of the invention [0005] The purpose of the present invention is to provide...

Claims

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Application Information

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IPC IPC(8): G01N33/543G01N33/535G01N33/68G01N21/76
CPCG01N33/54306G01N33/535G01N33/54326G01N33/68G01N21/76
Inventor 周靖彭英丽
Owner 捷和泰(北京)生物科技有限公司
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