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Method for preparing latex reagent by using polystyrene binding peptide and single-chain antibody

A single-chain antibody and polystyrene microsphere technology, applied in chemical instruments and methods, carrier binding/fixed peptides, hybrid peptides, etc., can solve the problems of rare scFV, loss of antigen binding, etc., to reduce non-specific reactions, The effect of small steric hindrance and improving the stability of reagents

Inactive Publication Date: 2014-10-22
唐勇
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Problems solved by technology

[0007] Although scFV is a preferred material form for studying antigen-antibody reactions, it is difficult to directly and stably immobilize the surface of the hydrophobic polystyrene matrix in its own state, and even if it is immobilized on the surface, it almost loses the ability of antigen binding. , so the application of scFV in diagnostic reagents is very rare

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  • Method for preparing latex reagent by using polystyrene binding peptide and single-chain antibody
  • Method for preparing latex reagent by using polystyrene binding peptide and single-chain antibody
  • Method for preparing latex reagent by using polystyrene binding peptide and single-chain antibody

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Embodiment Construction

[0022] The present invention will be further described below in conjunction with specific examples, and these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0023] 1. Acquisition of single-chain antibody gene sequence

[0024] RNA can be extracted from cell lines of high-affinity monoclonal antibodies screened by traditional hybridoma methods, and the coding sequences of the variable regions of the heavy chain and light chain can be obtained by PCR amplification with specific primers. It can also be screened from a constructed phage library targeting a specific antigen by displaying a single-chain antibody phage library. The heavy chain and light chain variable region coding sequences of the CRP single-chain antibody in this specific embodiment are amplified from monoclonal antibody cell lines, and the single-chain antibody is amplified with a hydrophilic flexible peptide (Gly4Ser)3 The variable r...

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Abstract

The invention provides a method for preparing a latex immunonephelometric assay kit with more specific performance, a more intact antibody stereostructure, higher affinity and fewer interference factors by orientedly immobilizing a single-chain antibody on the surface of a polystyrene microsphere through fusion expression of scFV and polystyrene binding peptide on the basis of the characteristic that the polystyrene binding peptide can specifically adhere on the surface of the polystyrene microsphere.

Description

technical field [0001] The invention relates to an immunoturbidimetric detection method, in particular to the development of a latex-enhanced immunoturbidimetric detection kit for single-chain antibody preparation mediated by the technology of directional immobilization of single-chain antibodies to the surface of polystyrene microspheres. Background technique [0002] The latex particle-enhanced turbidimetric immunoassay (PETIA) is a relatively stable and accurate homogeneous immune turbidimetric detection method for body fluid proteins that has emerged in recent years. Antibody, when the cross-linked antibody microspheres are combined with the antigen, they will quickly gather together in a short time, changing the astigmatism or light transmission performance of the reaction solution, and the astigmatism or light transmission performance (ie absorbance) of the reaction solution. The change has a strong correlation with the concentration of the tested antigen, and can refl...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/545G01N35/00C07K7/06C07K7/08C07K19/00C07K17/08
CPCG01N33/545G01N33/54313G01N33/54393
Inventor 唐勇
Owner 唐勇
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