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Technological process of continuously perfused culture of hybrid tumor and CHO cell to prepare antibody

A process method and hybridoma technology, applied in the field of bioengineering, can solve the problems of low adsorption efficiency and short life of monoclonal antibody, and achieve ideal effect, low cost and easy operation

Inactive Publication Date: 2002-03-13
陈志南
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  • Application Information

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Problems solved by technology

Streamliner Protein A is an affinity chromatography medium, so it can be used for one-step purification to achieve higher purity and recovery, but this medium has a shorter lifespan, and its price is nearly 16 times higher than that of streamline SP, and it is also suitable for Murine IgG subclass mAb adsorption is less efficient

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  • Technological process of continuously perfused culture of hybrid tumor and CHO cell to prepare antibody
  • Technological process of continuously perfused culture of hybrid tumor and CHO cell to prepare antibody
  • Technological process of continuously perfused culture of hybrid tumor and CHO cell to prepare antibody

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Embodiment Construction

[0013] The target product of the present invention is a mouse-derived monoclonal antibody secreted by hybridoma cells (HAb18). 31 I-HAb18 F(ab') 2 Hepatocarcinoma monoclonal antibody injection has entered Phase II and Phase III clinical trials. Liver cancer monoclonal antibody HAb18 IgG1 and F(ab') 2 Fragment antibodies have high specificity and affinity for primary hepatocellular carcinoma. The preparation process and technical route of the present invention can be applied to pilot-scale research and large-scale production of monoclonal antibodies and humanized antibodies. The steps are as follows:

[0014] 1. Large-scale culture of hybridoma cells (HAb18):

[0015] 1. Preparation of seed cells

[0016] (1) Expansion of seed cells

[0017]HAB18 hybridoma cells were prepared by immunizing BABL / c mice with the cell suspension of fresh liver cancer surgical specimens and fused the splenocytes with mouse myeloma SP / 20 cells. Established in August 1987, it has been stably s...

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Abstract

The technological process includes inoculation of cell to carrier in fixed bed; continuous perfused culture; detection; regulation and analysis of cell metabolism, growth rate and product synthesis balance; and product recovering and purification. The regulation and analysis of cell metabolism, growth rate and product synthesis balance includes the analysis of glucose, lactic acid, pH and other indexes, adding corresponding amino acid before the cell decline phase, decreasing glucose and blood serum concentration, etc. to realize high density and high yield culture. The product recovering andpurification inclues utilizing Streamline SP medium to recover antibody in supernatant, utilizing DEAE-Sepharose FF anionic exchanger to purify product, freeze drying.

Description

technical field [0001] The invention belongs to bioengineering technology, large-scale cultivation of animal cells (herein referring to hybridomas and CHO cells) and a preparation process for protein production by animal cells Background technique [0002] The wide application of engineered antibodies in basic medical research, clinical diagnosis and treatment, and immune prevention has greatly promoted the process of industrialization. The original monoclonal antibody production method is to inject hybridoma cells into the peritoneal cavity of mice, collect ascites and purify to obtain monoclonal antibodies. This production method is mainly used in laboratory research and preclinical research, and cannot be industrialized, and the pollution of rodent animal infection pollutants is inevitable, so it is difficult to meet the needs of clinical application. At present, the main method of industrial production of monoclonal antibodies is to use microcarrier fermenters, hollow f...

Claims

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Application Information

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IPC IPC(8): C12N5/12C12P21/00
Inventor 米力李玲冯强余晓玲
Owner 陈志南
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