Preparation method for recombinant rabbit monoclonal antibody

A monoclonal antibody and antibody technology, applied in chemical instruments and methods, peptides, specific peptides, etc., can solve the problems of low yield, single epitope, low affinity of monoclonal antibody and monoclonal antibody, and achieve high production efficiency, Easy operation, high affinity effect

Inactive Publication Date: 2020-06-02
安徽环球基因科技有限公司
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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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  • Preparation method for recombinant rabbit monoclonal antibody

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Embodiment 1

[0026] A preparation method of recombinant rabbit monoclonal antibody, the specific steps are as follows:

[0027] Step S1: Cut off the hair on the rabbit’s back, wipe the rabbit’s back with alcohol, add 1 mg of recombinant protein and an equal volume of Freund’s complete adjuvant into a beaker, stir until fully emulsified, and prepare the first mixed solution, and the second A mixed solution was injected subcutaneously on the back of the rabbit, and after 21 days of immune enhancement, 0.5 mg of recombinant protein and an equal volume of Freund's complete adjuvant were added to a beaker, and stirred until fully emulsified to prepare the second mixed solution. The solution was injected subcutaneously into the back of the rabbit, and the immune enhancement was carried out for 21 days, and the immune enhanced rabbit was obtained;

[0028] Step S2: Collect 10ml of blood from the ear vein of the immune-enhanced rabbit obtained in step S1, and use rabbit lymphocyte separation mediu...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/00
CPCC07K16/005C07K2317/20C07K2317/33C07K2317/76C07K2317/92
Inventor 雍金贵缪连军张磊
Owner 安徽环球基因科技有限公司
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