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High performance quick purifying method for preparing piecewise antibody

A technology of antibodies and fragments, which is applied in the field of bioengineering to achieve the effects of saving time, maintaining activity, and small differences in results

Active Publication Date: 2006-01-04
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0015] Sullivan; John B, Russell; Findlay E. In the patent related to antibody purification process (United States Patent, No 4,849,352) [4], it is described that papain-cleaved Fab fragment antibody is separated by polyphenylacrylamide gel affinity chromatography and pepsin-cleaved F(ab') 2 Antibodies, the conditions and methods of the purification process are not specifically described

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  • High performance quick purifying method for preparing piecewise antibody
  • High performance quick purifying method for preparing piecewise antibody
  • High performance quick purifying method for preparing piecewise antibody

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Embodiment Construction

[0063] The present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments given by the inventor.

[0064] According to the technical scheme of the present invention, the applicant uses the monoclonal antibody secreted by hybridoma cells to carry out F(ab') 2 Preparation of fragment antibodies. The hybridoma cell stably secretes and expresses a monoclonal antibody against human liver cancer-related antigen (preserved by the General Microorganism Collection Center of China Committee for Culture Collection of Microorganisms, deposit number: CGMCC No 0426).

[0065] Hybridoma cells were cultured in large-scale high-density suspension in a mechanically stirred bioreactor; the antibody IgG in the large-scale cell culture medium in the early stage was adsorbed by expanded column bed cation chromatography and concentrated by anion chromatography. Elution was carried out by changing the pH of the buffer system. Antib...

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Abstract

The invention discloses a method for high-effectively quickly purifying and preparing fragment antibody from animal cell scale culture. Said method adopts the processes of dilatant column bed adsorption, anion chromatography concentration and hydrophobic chromatography purification, and includes the following steps: using dilatant column bed cation chromatography to adsorb antibody IgG from large-scale cell culture, changing pH value of buffer system to make elution, making collected elution peak be directly andergone the process of anion chromatography, using pepsinum to directly enzyme-out the fully-concentrated and purified antibody IgG to obtain fragment antibody, making hydrophobic chromatography and purification so as to obtain final fragment antibody whose purity is up to above 97%, and its whole purification process can be completed within 12 hr. Said invented method can be substituted for traditional ammonium sulfate precipitation, concentration method, and has the characteristics of high efficiency, rapid speed, high purity and stable operation and result, etc.

Description

technical field [0001] The invention belongs to the protein purification technology in the technical field of bioengineering, and mainly relates to the purification and preparation process of antibodies including monoclonal antibodies and small molecule fragment antibodies, humanized or human-mouse chimeric antibodies and fragment antibodies. In particular, it relates to a method for efficiently and quickly purifying and preparing fragment antibodies, which is suitable for the production and preparation of therapeutic antibody drugs. Background technique [0002] Antibody therapy has become an important means of biological treatment of tumors, epidemics and infectious diseases today. As of February 2004, the US FDA has approved 16 antibody therapeutic drugs for marketing, and the market sales have exceeded 2×10 9 dollars. Antibody drugs have become a large class of new therapeutic agents in the international pharmaceutical market due to their high specificity, effectivenes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/16C07K16/18C12P21/06
Inventor 米力唐浩余晓玲谢丽
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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