Tag peptide capable of realizing affinity binding with polystyrene and method for preparing enzyme-linked immuno solid phase antigen with same

Abstract

The invention discloses a tag peptide capable of realizing affinity binding with polystyrene and a method for preparing enzyme-linked immuno solid phase antigen with the same. The tag peptide capable of realizing affinity binding with polystyrene has the amino acid sequence of Phe-Lys-Phe-Trp-Pro-Tyr-Glu-His-Val-Ile-Arg-Gly and the coded nucleotide sequence of TTCAAA TTC TGG CCA TAC GAA CAT GTA ATA CGG GGG. The tag peptide has the advantages that Lig2 tag peptide has strong affinity binding capability with a polystyrene carrier, and can guide a fusion antigen in a unified and orderly mode to be fixed on the surface of the polystyrene, thereby avoiding the phenomena of comformational change or site shielding because an antibody binding site directly contacts the rigid surface of the carrier, maintaining antibody binding activity and good spatial orientation to the greatest extent, and improving the enzyme-linked immuno detection sensitivity by 2.5-3 times; the tag peptide can be used for diagnosis detection technology, such as enzyme-linked immuno, biosensors, microarray chips and the like which take the polystyrene as a fixed carrier, and improves detection sensitivity.

Description

technical field [0001] The invention relates to a label peptide capable of affinity binding with polystyrene, and a method for improving the immobilization effect of antigen on the surface of polystyrene carrier by using it, so as to improve the detection sensitivity of ELISA. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) detects antibodies by immobilizing specific antigens on the surface of polystyrene carriers to form solid-phase antigens, which are then combined with the corresponding antibodies in the samples to be tested (such as serum). Its detection, this technology has been widely used in various fields of medical testing, and has become a routine method for clinical diagnosis [Butler JE, Journal ofimmunoassay & immunochemistry. 2000, 21: 165-209]. Among them, the immobilization of the antigen is the key link: the activity of the antigen is closely related to its three-dimensional structure and site orientation, and the antigen immobilized o...

AI Technical Summary

Problems solved by technology

Covalent immobilization is relatively stable, but there are many functional groups such as amino groups on the antigen molecule, which leads to the inability of directional binding of the antigen. In addition, the antigen immobilized on the surface of the carrier through covalent bonds is often denatured due to structural changes [Zhang Aijuan et al., Progress in Chemistry. 2009 , 21:1408-1417]

Whether it relies on non-covalent bonds or covalent bonds to bind to the carrier, this combination is random for antigen molecules, and the molecules are arranged in a disordered state on the surface of the carrier, such as figure 1 As shown, only the solid-phase antigen with the exposed antibody binding site can bind to the target antibody, and some solid-phase antigen molecules cannot recognize the corresponding antibody because the binding site is covered, and cannot produce effective immunity in the subsequent detection of ELISA. response, it is difficult to meet the requirement of high sensitivity [Peluso P et al, Analytical Biochemistry.2003, 312:113-124]

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