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Efficient expression vector of antibody and preparation method for efficient expression vector

A high-efficiency expression and vector technology, applied in the field of genetic engineering, can solve the problem of unbalanced expression of antibody heavy chain and light chain, and achieve the effect of avoiding unbalanced expression, high expression rate and improving effectiveness.

Active Publication Date: 2012-07-11
广东安普泽生物医药股份有限公司
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Problems solved by technology

[0007] In view of this, the technical problem to be solved by the present invention is to provide an antibody high-efficiency expression vector, which can simultaneously express the heavy chain and light chain of the antibody, avoiding the existing The problem of unbalanced expression of antibody heavy chain and light chain in the technology, while increasing the expression rate of antibody protein, can improve the effectiveness of subsequent screening of monoclonal antibody cell lines

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  • Efficient expression vector of antibody and preparation method for efficient expression vector
  • Efficient expression vector of antibody and preparation method for efficient expression vector
  • Efficient expression vector of antibody and preparation method for efficient expression vector

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Embodiment Construction

[0044] In order to make the present invention easier to understand, the present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0045] For the experimental methods not indicated in the following examples, the specific conditions are generally in accordance with conventional conditions, such as those described in the Molecular Cloning Implementation Manual of Sambrook et al.

[0046] 1. Construction of the high expression plasmid of the present invention

[0047] In the following plasmid construction process, the PCR products were purified by Qiagen kit MinElute Gel Extraction Kit (Cat. No. 28604) and then cleaved. If it is necessary to replace the endonuclease buffer during digestion, use the kit QIAquick PCR Purification Kit (Cat. No. 28104) from Qiagen. Except that endonuclease Bst...

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Abstract

The invention discloses an efficient expression vector of an antibody. The efficient expression vector contains resistance genes and two target gene binding sites for binding the genes, the two target gene binding sites can be respectively combined with heavy chain genes and light chain genes of antibody molecules, the two target gene binding sites are respectively positioned in two expression units in the efficient expression vector and respectively form a heavy chain expression unit and a light chain expression unit of the antibody, and each expression unit also comprises a strong expression inductivity promoter positioned on the upstream of each target gene binding site. The efficient expression vector can simultaneously express the heavy chain and the light chain of the antibody, solve the problem about imbalanced expression of the heavy chain and the light chain of the antibody in the prior art, increase the expression rate of antibody protein and improve the screening effectiveness of subsequent monoclonal antibody cell strains.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a high-efficiency expression vector for animal cells and a construction method thereof. Background technique [0002] Antibody is an immunoglobulin molecule with special amino acid sequence induced by antigen, synthesized and secreted by lymphocytes (plasma cells). Antibodies can bind to antigens such as bacteria, viruses or toxins to neutralize or remove harmful substances through several different mechanisms. Antibodies have been studied as therapeutic agents for a century (Drews J. Drug discovery: a history perspective. Science, 2000, 287(5460): 1960-1964). Antibody drugs are currently a particularly active field in biotechnology pharmaceuticals, playing an important role in the treatment of certain diseases. [0003] In 1975, Kohler and Milstein reported the preparation of monoclonal antibody (monoclonal antibody, MAb) by B lymphocyte hybridoma technology. Mono...

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Application Information

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IPC IPC(8): C12N15/85C12N15/66
Inventor 陈倩怡王笑非
Owner 广东安普泽生物医药股份有限公司
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