Method and kit for identifying sensitivity of antibody and clone cell strain

Abstract

The invention discloses a method for identifying sensitivity of an antibody as well as quality of a clone cell strain in the preparation process of the antibody. The method comprises the following steps: acquiring a solid-phase carrier, cells and the antibody, adsorbing the antibody on the solid-phase carrier, incubating the cells and the antibody adsorbed on the solid-phase carrier, keeping cells that are combined with the antibody, dyeing and counting the cells combined with the antibody, and identifying the sensitivity of the antibody and the quality of the clone cell strain according to the cell counting result. The invention further provides a kit applicable to the method. The method intuitively reflects the level of combination of the antibody and a native conformation antigen, and the identifying method is simple, quick and intuitive.

Description

technical field [0001] The invention relates to the technical field of antibodies. More specifically, the present invention relates to a method for identifying antibody sensitivity, and a method for identifying positive clonal cell lines secreting target antibodies during antibody preparation. Background technique [0002] Antibodies are a type of globulin with immune function secreted by B cells that proliferate and differentiate into plasma cells. They exist in body fluids and mediate humoral immunity. Antibodies can specifically bind to corresponding antigens (such as pathogens), and exert immune effects with the participation of other immune molecules and cells. According to different properties, antibodies can be divided into monoclonal antibodies (hereinafter referred to as monoclonal antibodies) and polyclonal antibodies (hereinafter referred to as polyclonal antibodies). Monoclonal antibodies have the advantages of highly uniform physical and chemical properties, s...

AI Technical Summary

Problems solved by technology

[0005] Immunofluorescence test includes direct immunofluorescence test and indirect immunofluorescence test, in which indirect immunofluorescence test needs to be completed with the help of secondary antibody. Long, cumbersome operation steps, and expensive experimental equipment, such as fluorescence microscope, flow cytometer

[0006] CN1660907A discloses three identification methods of monoclonal antibodies, including indirect immunofluorescence, dot immunoblotting and gold-labeled immunoelectron microscopy, in which both indirect immunofluorescence and gold-labeled immunoelectron microscopy require expensive equipment to cooperate with the judgment of the results. The dot immunoblotting method uses nitrocellulose membrane as a solid phase carrier for antigen-antibody reaction, and needs to obtain cell membrane protein lysate for adsorption. Since the protein in the cell membrane protein lysate is denatured, the identification result cannot reflect the monoclonal antibody and natural conformational protein sensitivity

[0007] CN1718588A discloses a method of monoclonal antibody for malignant tumor detection, which is realized by ELISA technology and immunoblotting technology, ELISA test technology uses recombinant protein as antigen for detection, and Western blotting test technology uses denatured protein as antigen Assays performed, neither test technique truly reflects the sensitivity of mAbs to native conformational proteins

[0008] CN101891806A discloses a screening method for the quality of monoclonal antibodies, which is realized by using indirect immunofluorescence technology. The judgment of indirect immunofluorescence test requires an expensive fluorescence microscope, and the economic affordability of general small clinical and scientific research institutions is limited. This method Not conducive to popularization

This method generally uses recombinant proteins as antigens, and the screened monoclonal antibodies are often not suitable for the binding reaction of natural conformational proteins, resulting in screening failure and waste of manpower and material resources

[0010] CN1693461A discloses the screening method of positive clonal cell strains in the preparation process of monoclonal antibody, is to use indirect enzyme-linked immunosorbent assay (ELISA) to carry out, this method generally is to use recombinant protein as antigen to screen, but because recombinant protein and natural conformation There are certain differences in proteins, so the positive clonal cell lines screened by ELISA cannot truly reflect the sensitivity of monoclonal antibodies and natural conformational proteins

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