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Antibody production method

A production method and antibody technology, applied in chemical instruments and methods, peptides, specific peptides, etc., can solve the problems of limited adjustment range of charge variants, poor versatility, and no significant increase in antibody yield, to increase antibody yield and reduce acidity. Peak content, the effect of improving charge heterogeneity

Inactive Publication Date: 2018-03-16
上海泰因生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have a limited adjustment range for charge variants, and their versatility is poor, and at the same time, they do not significantly improve the production of antibodies

Method used

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Embodiment 1

[0044] The antibody production process of this embodiment is as follows:

[0045] (a) Add basal culture medium to the cell culture flask, inoculate the GS-CHO antibody-expressing cell line, and carry out routine culture at 37°C after inoculation;

[0046] (b) After culturing for a certain period of time (the 4th day), the culture temperature was lowered to 34°C; at the same time, on the 4th day, the following strategy was used to add the culture solution containing the protein hydrolyzate until the end of the culture:

[0047] Once every two days, the volume of the fed-batch culture solution added each time is 5%, 2%, 5%, 2%, 5% of the culture volume;

[0048] c) Stop the cell culture when the cell viability drops to 50% or the cell density is lower than 1 / 3 of the highest cell density, and harvest the supernatant.

[0049] in,

[0050] The basic culture medium is Dynamis;

[0051] Fed-batch medium is Feed C;

[0052]The inoculated antibody expression cell lines were GS-CH...

Embodiment 2

[0062] The present embodiment adopts the method identical with embodiment 1 to cultivate, wherein,

[0063] The present embodiment selects 6 kinds of basic culture fluids as Dynamis, T54, Cellvento TM CHO-210, Cellvento TM CHO-200, ApmliCHO CD, OPM-CD11V;

[0064] Fed-batch medium is Feed C;

[0065] The protein hydrolyzate is soybean protein hydrolyzate, and the added concentration is 100g / L;

[0066] In this embodiment, the fed-batch culture solution without protein hydrolyzate was also used as a control group;

[0067] The inoculated cell line is GS-CHO cell antibody expression strain 2, and the cell inoculation density of the antibody expression cell line is (0.5*10 6 cells / ml);

[0068] The culture product was tested and the following results were obtained:

[0069] See Figure 4-Figure 6 , where, compared to the control group,

[0070] Using the culture medium Dynamis, the acidic peak content of the antibody decreased from 23.4% to 19.6%, the main peak content i...

Embodiment 3

[0078] The present embodiment adopts the method identical with embodiment 1 to cultivate, wherein,

[0079] The basal culture medium selected in this embodiment is Dynamis;

[0080] Fed-batch medium is Feed C;

[0081] The protein hydrolyzate is corn protein hydrolyzate, and the added concentration is 20g / L, 40g / L, 60g / L, 100g / L;

[0082] In this embodiment, the fed-batch culture solution without protein hydrolyzate was also used as a control group;

[0083] The inoculated cell line is GS-CHO antibody-expressing cell line 2, and the cell inoculation density of the antibody-expressing cell line is (0.5*10 6 cells / ml).

[0084] The culture product was tested and the following results were obtained:

[0085] See Figure 7-Figure 9 , where, compared to the control group,

[0086] When the concentration of protein hydrolyzate was 20g / L, the acidic peak content of the antibody was 19.8%, the main peak content was 72.1%, and the yield was 1.53g / L;

[0087] When the concentrati...

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Abstract

The invention discloses an antibody production method. The method starts from the metabolic process of cell culture, a protein hydrolysate is added into a fed-batch culture solution to achieve adequate nutrition and an inhibition and prevention effect on modification and degradation of partial charge heterogeneity in the expression process, thereby stably and effectively improving the charge heterogeneity of the antibody product and increasing the antibody expression. The method can effectively solve the charge heterogeneity problem in antibody production, and can maintain or improve the yield.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a production method for improving the charge heterogeneity of antibodies by adding protein hydrolyzate, and the production method can maintain or increase the yield at the same time. Background technique [0002] At present, monoclonal antibody drugs are rapidly developing in the drug market due to their specific targeting and low toxicity. In 2016, the global monoclonal antibody market sales reached 100 billion U.S. dollars, of which the "King of Medicine" adalimumab sales reached 16 billion U.S. dollars. [0003] Monoclonal antibody drugs are usually expressed by mammalian cells due to their large molecular weight and complex advanced structural modifications, among which CHO cells are the most mainstream host cells for expressing monoclonal antibodies. [0004] However, due to the influence of the complex culture environment during the production process of antibodies, some ...

Claims

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Application Information

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IPC IPC(8): C12P21/00
CPCC07K16/00C07K2317/14
Inventor 秦金燕郑琛齐念民庄超钱慧傅强伍翔
Owner 上海泰因生物技术有限公司
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