A kind of preparation method of enzyme labeling antigen antibody

Abstract

The invention discloses a preparation method of enzyme labeled antigens and antibodies. Firstly, 2-hydroxyphenylacetic acid in HRP (horse radish peroxidase) molecules is oxidized under the oxidability of sodium periodate to form aldehyde groups, a freeze-drying protective agent is added to an enzyme solution for freeze-drying treatment after micromolecular substances are removed through dialysis, and then the solution is left for standby use; during antigen and antibody labeling, the antigens and the antibodies are subjected to alkali treatment firstly, primary amino groups are exposed, aldehyde HRP molecules subjected to freeze-drying treatment are added, a crosslinking reaction is generated and lasts for 1-1.5 h, an anti-freezing protective agent is added after the reaction ends, and the enzyme labeled antigens and antibodies can be stored for a long time at the low-temperature condition of subzero 20 DEG C. According to the preparation method, a protein crosslinking technique is combined with a freeze-drying technique, a traditional sodium periodate method is improved and optimized, the experimental period of the enzyme labeled antigens and antibodies is greatly shortened, the crosslinking efficiency of HRP and the antigens as well as the antibodies is improved, and the reactivity of enzyme labeled conjugates is improved.

Description

technical field [0001] The invention relates to the technical field of immunological detection, in particular to a method for preparing an enzyme-labeled antigen-antibody. Background technique [0002] Immunological detection technology has the advantages of fast detection speed, low cost, simple and portable equipment, high sensitivity and strong selectivity, and can be used for on-site inspection. It combines the amplification effect of enzyme-catalyzed reaction with the high specificity and specificity of antigen-antibody affinity reaction, and uses enzyme-labeled antigen or antibody as the key reagent for immunoassay, so it has high sensitivity. Enzymes used to label antigens or antibodies must have the following characteristics: high activity and sensitivity; stable at room temperature; easy visualization of reaction products; and commercial production. Horseradish peroxidase (HRP), alkaline phosphatase, glucose oxidase, etc. are currently widely used, among which HRP ...

AI Technical Summary

Problems solved by technology

[0004] The traditional sodium periodate method has some disadvantages when labeling some antigens and antibodies, such as long experimental period, low cross-linking efficiency, poor reactivity of enzyme-labeled antigens and antibodies, etc., which limits its use to a certain extent. application