Biotinylation bovine serum albumin and streptavidin enzyme-labeled reaction plate and preparation method thereof

Abstract

The invention discloses a biotinylation bovine serum albumin and streptavidin enzyme-labeled reaction plate and a preparation method thereof, belonging to the field of labeling immunoassay in laboratory medicine. The reaction plate comprises a polystyrene enzyme-labeled plate; each micropore is respectively coated with the biotinylation bovine serum albumin and the streptavidin; and a biotinylation antibody is stored in a liquid phase to serve as a reaction reagent for later use. When in detection, the biotinylation antibody and an antigen to be detected are simultaneously added on the enzyme-labeled reaction plate; the biotinylation antibody and the antigen to be detected are simultaneously reacted; and biotin molecules and the streptavidin are combined and fixed on the surface of solid phase materials. According to the invention, the antigen to be detected and the biotinylation antibody are placed in the liquid phase so as to improve reaction rate, widen detection range of ferritin, and reduce hook effects in double antibody sandwich experiment. Indexes such as reaction sensitivity, accuracy, precision and the like of the enzyme label satisfy the technical requirements; the usage ratio of the coated antibody is improved; the usage amount of the antibody is reduced; and the cost of a reagent box is greatly reduced.

Description

technical field [0001] The invention relates to enzyme-linked immunoassay technology, in particular to an enzyme-labeled reaction plate coated with biotinylated bovine serum albumin and streptavidin and a preparation method thereof. Background technique [0002] Enzyme linked immunosorbent assay (ELISA) uses an enzyme-labeled antibody (or antigen) and an enzyme-labeled reaction plate coated with an antibody (antigen) as the main reagents, combining the specificity of the antigen-antibody reaction with the enzyme-catalyzed substrate. A marker immunoassay technique that combines high-efficiency materials. The main technology types of ELISA include: double antibody sandwich method, indirect method, competition method, capture method, etc. The double-antibody sandwich method is one of the commonly used techniques and can be used to detect trace substances in the body. [0003] Enzyme-linked immunosorbent assay is a heterogeneous immunoassay, and it is necessary to separate the...

AI Technical Summary

Problems solved by technology

[0007] First, the antibody molecules are adsorbed on the surface of the solid-phase material. Since the antigen-antibody reaction depends on the spatial conformation, the conformational change caused by the solid-phase of the antibody will inevitably affect the utilization efficiency of the antibody, resulting in a decrease in the affinity between the antigen and the antibody.

[0008] Second, the small surface area in the microwells of the microenzyme plate leads to a limited number of coated antibody molecules, and the reduction in utilization rate caused by solid-phase conversion will make the number of antibody molecules unable to meet the needs of the antigen to be tested, affecting the number of antibody molecules. The detection range and detection time of the test method

[0009] Third, the solid-phase antibody molecules are in a static state, so the probability of collision between the antibody on the solid phase and the antigen in the liquid phase is always smaller than that of the liquid phase reaction, which will inevitably increase the time required for the antigen-antibody reaction to reach equilibrium

At the same time, in the one-step method, the antigen to be tested and the enzyme-labeled antibody need to be added at the same time. The reaction rate of the antigen to be tested and the capture antibody is lower than the reaction rate of the antigen to be tested and the enzyme-labeled antibody, resulting in the occurrence of "groove effect" and false negative results.

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