A kind of micropolymer-hrp-nanometer antibody complex and preparation method thereof

Abstract

The invention belongs to the technical field of immune analysis, and in particular relates to a micropolymer-HRP-nanometer antibody complex and a preparation method thereof. The micropolymer, HRP and Nanobody in the micropolymer-HRP-Nanobody complex provided by the present invention are all obtained through different chemical modifications, wherein the amino acid sequence of the Nanobody is shown in SEQ ID NO.1. The complex provided by the present invention greatly reduces the volume of the immunohistochemical secondary antibody, improves its molecular movement ability and penetration ability, improves its staining intensity and staining sensitivity, and at the same time improves the batch-to-batch ratio of the immunohistochemical secondary antibody. stability.

Description

A kind of micropolymer-HRP-nanobody complex and preparation method thereof technical field The invention belongs to the technical field of immunoassay, be specifically related to a kind of micropolymer-HRP-nanobody composite substances and methods for their preparation. Background technique Immunohistochemistry (Immunohistochemistry) utilizes the specific binding principle of antigen and antibody and Special labeling technology, through chemical reaction, the secondary antibody labeled with fluorescein, enzyme, metal ion, isotope is in the display agent. Under the action of color development, it is a technology for localization, qualitative and relative quantitative detection of specific antigens in tissue cells. waived Immunohistochemical (IHC) staining is of great significance for tumor determination and medication guidance. Immunohistochemical analysis is very important for tumor molecular typing. Accurate tumor typing detection is conducive to guiding patients...

AI Technical Summary

Problems solved by technology

However, this method is not perfect, and its main technical defect is: the method uses a dextran polymer backbone with a molecular weight ≥ 500,000, which connects 4-70 enzyme molecules and 1-10 antibody molecules, and its structure is loose and fluid. The large dynamic volume creates a great steric hindrance, which makes the polymer have low penetration of the nuclear membrane

However, this technique has two disadvantages: one is that the highly toxic dimethyl sulfoxide is used as the coupling reagent many times in the preparation process, which is harmful to human health and the environment; the other is that the secondary antibody used in the preparation is mostly Antibody and polyclonal antibody need to be obtained by immunizing animals, the production cycle is long, the process is complicated, the production controllability between batches is poor, and the polyclonal antibody has strong cross-reaction with other antibodies, which may easily lead to non-specific background staining

[0009] The Chinese invention patent with the application number 202011221677.X provides a small polymer enzyme-antibody fragment with a dendritic polymer as the backbone, which makes the enzyme connected to the backbone more compact and reduces the space occupation. Then it is coupled with a half antibody to halve the mass and volume of the antibody, and finally a heterobifunctional coupling reagent is used to connect the small polymerase complex to a specific site of the antibody fragment; although the final antibody fragment has improved Tissue penetration ability, the advantage of effectively reducing the staining background, but this small polymer enzyme-antibody fragment is likely to affect the staining sensitivity and destroy the stability of immunohistochemical secondary antibody batches

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