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Method for producing anti CD20 antibody

A CCTCCNO.C201158, vitamin technology, applied in the field of biopharmaceuticals, can solve the problems of large differences in different batches, high cost, microbial contamination, etc., and achieve the effect of increasing antibody expression and promoting growth

Active Publication Date: 2015-04-15
LUNAN PHARMA GROUP CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, on the one hand, animal serum will provide substances such as hormones, antibiotics, and trace elements for cell growth; The difference is large and the cost is high

Method used

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  • Method for producing anti CD20 antibody
  • Method for producing anti CD20 antibody

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The medium was prepared according to the components and contents described in Table 2, and the medium was sterilized by filtration with a 0.22 μm bacterial filter membrane before being used for production. A cryopreserved CHO cell was taken out from the liquid nitrogen of the production cell bank, revived and inoculated into a 100 mL spinner bottle for culture. In the spinner bottle culture stage, the shaker speed was 85 rpm and the culture temperature was 37°C.

[0028] Amplify the spinner bottle step by step, after 100mL→250mL→500mL passage expansion, when the total number of cells reaches 1.0×10 8 One hour, inoculated into a 5.0L reactor for cultivation, the inoculum size was 0.3×10 6 cells / ml culture medium. During the fermentation process, the glucose concentration was maintained at 0.5-3g / L, and the dissolved oxygen was 30%. From the initial stage of culture to the logarithmic growth phase, control the culture temperature to 37°C, pH to 7.2, and stirring speed t...

Embodiment 2

[0033] The medium was prepared according to the components and contents described in Table 3, and the medium was sterilized by filtration with a 0.22 μm bacterial filter membrane before being used for production. A cryopreserved CHO cell was taken from the liquid nitrogen of the production cell bank, which is a cell with high expression of anti-CD20 antibody. After recovery, it was inoculated into a 100mL spinner bottle for culture. During the spinner bottle culture, the shaker speed was 75rpm and the culture temperature was 36°C.

[0034] Amplify the spinner bottle step by step, after 100mL→250mL→500mL passage expansion, when the total number of cells reaches 1.0×10 8 One hour, inoculated into a 5.0L reactor for cultivation, the inoculum size was 0.5×10 6 cells / ml culture medium. During the fermentation process, the glucose concentration was maintained at 0.5-3g / L, and the dissolved oxygen was 50%. From the initial stage of culture to the logarithmic growth phase, control t...

Embodiment 3

[0040] The medium was prepared according to the components and contents described in Table 4, and the medium was sterilized by a 0.22 μm filter membrane before being used for production. Take out a cryopreserved CHO cell from the liquid nitrogen of the production cell bank, which is a high-expression anti-CD20 antibody cell. After recovery and expansion, it is inoculated into a 100ml spinner bottle for culture. During the spinner bottle culture, the shaker speed is 75rpm and the culture temperature is 37. ℃.

[0041] Amplify the spinner bottle step by step, after 100mL→250mL→500mL passage expansion, when the total number of cells reaches 1.0×10 8 One hour, inoculated into a 5.0L reactor for cultivation, the inoculum size was 0.5×10 6 cells / ml culture medium. During the fermentation process, the glucose concentration was maintained at 0.5-3g / L, and the dissolved oxygen was 60%. From the initial stage of culture to the logarithmic growth phase, control the culture temperature...

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Abstract

The invention discloses a method for culturing animal cells for producing anti CD20 antibody, and specifically discloses inoculation quantity, medium formula and various culture process control parameters. The medium is a serum-free medium, so that no side effect is caused by serum in the process of cell culture, and antibody extraction and purification are further facilitated. By using the method for producing the anti CD20 antibody, differences between different batches are small, antibody expression quantity is high, production cost is low, and safety is good.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method for producing an anti-CD20 antibody. Background technique [0002] Non-Hodgkin's lymphoma (NHL) is one of the most common clinical malignant tumors, about 85% of which are derived from B cells, and its morbidity and mortality have ranked fifth among malignant tumors. NHL has a high effective rate for traditional treatment methods such as chemotherapy, radiotherapy, and immune regulation, but the recurrence rate is also high, the remission rate of retreatment is low, and the toxicity is large, and the patient's tolerance is poor. In the past ten years, with the development of antibody drugs, anti-CD20 monoclonal antibody preparations represented by MabThera have greatly improved the clinical efficacy of lymphoma. [0003] At present, in the method of culturing animal cells to prepare antibodies, the medium containing more than 10% serum is mostly used to make the cells ...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12P21/00C12R1/91
Inventor 赵志全赵丽丽程凡亮刘增田张继环
Owner LUNAN PHARMA GROUP CORPORATION
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