Anti-CD20 monoclonal antibody, preparation method and application thereof

Abstract

The invention relates to an anti-CD20 monoclonal antibody, a preparation method and an application of the anti-CD20 monoclonal antibody; Chinese hamster ovary cells CHODG44 are adopted to prepare the anti-CD20 monoclonal antibody consistent to rituximab, thereby establishing a large-scale high-expression production technology for 300L of eukaryocytes, wherein the protein expression quantity is more than 1.2g / L and the protein purification yield is improved to more than 60%. The anti-CD20 monoclonal antibody is safe, effective, relatively strong in practicability, low in cost, convenient and quick, and the preparation technology of the anti-CD20 monoclonal antibody is simple and convenient, easy to operate and use and obvious in curative effect, thereby providing a new source of medicines for the B cell induced malignant tumors, particularly the non-hodgkin lymphoma, autoimmune disease, renal failure, hepatitis B, hepatitis C and diabetes, etc.; the anti-CD20 monoclonal antibody is applied to the large-scale production and commercial application in the industries such as medicine, biotechnology and the like and has a good application prospect, remarkable social benefit and economic benefit.

Description

technical field [0001] The present invention relates to the fields of medicine, biotechnology, etc., specifically to a monoclonal antibody, antibody humanization, and other biotechnology and medical technology fields, as well as its preparation method and use, and more specifically to a monoclonal antibody The cloned antibody and its preparation method and use, more specifically relate to an anti-CD20 monoclonal antibody (abbreviation: anti-CD20 monoclonal antibody) and its preparation method and use. Background technique [0002] (1) Research overview of anti-CD20 monoclonal antibody [0003] 1 Overview [0004] The mortality rate of tumors ranks first in the mortality rate of various diseases in the world, and malignant tumors have become a killer that seriously affects human health and life. According to incomplete statistics, in China, there are more than 3 million new patients with malignant tumors every year, of which 1.3 million people die of malignant tumors. In t...

AI Technical Summary

Problems solved by technology

But at the same time, there are still many insurmountable shortcomings in the prokaryotic expression system: for example, the commonly used expression system cannot regulate the expression time and expression level, the continuous expression of some genes may have toxic effects on the host cells, and overexpression may lead to abnormal Physiological reactions, the target protein is often expressed in the form of inclusion bodies, resulting in difficulties in product purification; and the post-translational processing and modification system of the prokaryotic expression system is imperfect, and the biological activity of the expressed product is low

[0053] (2) Due to the limitations of animal serum extraction, it is extremely expensive to apply it to the culture medium

[0054] (3) The limitation of animal serum extraction also brings difficulties to the standardization of cell culture. At the same time, there is also the problem of difficult separation and purification of cell culture expression products, which have potential cytotoxicity and increase the difficulty of large-scale culture of cells (Even MS ,Sandusky C B,Barnard N D.Serum free hydraulicoma culture:ethical,scientific and safety considerations[J].Trendsin Biotechnology,2006,24(3):105-108.)

[0063] ① Cells are susceptible to certain mechanical and chemical factors in serum-free medium, and the preservation and application of the medium are not as convenient as traditional synthetic medium

[0064] ② Higher cost

[0112] However, through literature search, etc., so far, no new anti-CD20 products and their preparation methods and uses have been found.

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