Preparation method for recombinant rabbit monoclonal antibody

Abstract

The invention discloses a preparation method for a recombinant rabbit monoclonal antibody. The method utilizes immune-raised rabbit B cells for isolation, phage display and other technologies, antibody gene fragments are amplified through specific primers, an antibody library is constructed, an antigen is used for panning to obtain positive clones, sequence identification is performed to obtain heavy and light chain sequences, in-vitro recombinant expression is performed, and the antibody is measured; compared with a traditional method for preparing a monoclonal antibody, the preparation method for the monoclonal antibody provided by the invention is higher in efficiency of preparing the monoclonal antibody and more convenient to operate; and compared with a mouse monoclonal antibody, themonoclonal antibody screened by using an immune system of an experimental rabbit and capable of recognizing a specific antigen has the prominent characteristics of high affinity and more abundant recognized epitopes, and can recognize antigens having no immunogenicity in mice.

Description

technical field [0001] The invention belongs to the field of monoclonal antibody preparation, and in particular relates to a preparation method of recombinant rabbit monoclonal antibody. Background technique [0002] Antibody refers to a protective protein produced by the body due to antigen stimulation. It is a large Y protein secreted by plasma cells (effector B cells) and used by the immune system to identify and neutralize foreign substances such as bacteria and viruses. Antibodies play an important role in disease diagnosis, immune prevention and basic research. Antibodies are divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies have broad prospects in medical treatment and are used to treat tumors, self Various diseases such as immune diseases, infectious diseases and transplant rejection; [0003] Chinese invention patent CN1763217A discloses a preparation method and application of a monoclonal antibody. In this invention, sterilized St...

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Problems solved by technology

[0003] Chinese invention patent CN1763217A discloses a preparation method and application of a monoclonal antibody. In this invention, sterilized Staphylococcus aureus is used to immunize BALB / c mice, and then the splenocytes of the immunized mice are fused with Sp2 / 0 myeloma cells. The hybridoma strain secreting anti-Staphylococcus aureus monoclonal antibody is obtained by screening and cloning, and the desired monoclonal antibody is obtained. The invention further obtains the monoclonal antibody through hybridoma cells. The preparation efficiency is low and the yield is not high. The monoclonal antibody screened by the mouse immune system has a low overall affinity, and the recognized epitope is relatively single, and cannot recognize non-immunogenic antigens

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