Preparation method for monoclonal antibody

A monoclonal antibody, antibody technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc., can solve the problems of high experimental costs and conditions, inability to use, etc.
CN102776200AInactive Publication Date: 2012-11-14BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
Publication Date
2012-11-14
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a preparation method for a monoclonal antibody. According to the method, magnetic granular microballoon spheres of an immobilized antigen are added into B lymphocyte culture micropores, magnetic separation is carried out on the magnetic granular microballoon spheres so as to allow the spheres to enter into corresponding microplates, a single B lymphocyte which secretes a specific antibody is detected by using the method of immunochemiluminescence or immunofluorescence, a light chain variable region gene and a heavy chain variable region gene of the antibody are amplified by using the method of single-cell RT-PCR and nested RT-PCR and are recombined to expression plasmid containing a constant domain of the antibody, and a host cell is transfected to express the monoclonal antibody.
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Description

technical field

[0001] The invention relates to the technical fields of bioengineering and monoclonal antibody production. Specifically, the present invention relates to the detection and recognition of a single B cell that secretes an antigen-specific antibody, the antibody light and heavy chain variable region genes are amplified by RT-PCR and PCR methods, and constructed on a vector containing the constant region gene through homologous recombination , Transfected cells to prepare monoclonal antibodies. Background technique

[0002] Antibodies are produced by the body's immune system under the stimulation of antigens by the synthesis and secretion of plasma cells that proliferate and differentiate from B lymphocytes or memory cells. They are immunoglobulins that can specifically bind to corresponding antigens and are important in mediating humoral immunity. immune effector molecules. Antibodies have the functions of neutralizing viruses, activating the complement system...

Claims

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