Separation and purification method for cell culture mixture

A cell culture, separation and purification technology, applied in the field of separation and purification of biological products, can solve the problems of separation and purification of cell culture mixture, biological products with too many heterologous substances, complicated separation and purification process, etc., to achieve simple separation and purification steps and stable product quality. , the effect of a wide range of applications

Inactive Publication Date: 2011-09-21
BEIJING SKYWING TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Using existing technical means to separate and purify the cell culture mixture, due to the above problems, the separation and purification process is complicated, and the cell culture mixture cannot be separated and purified in large quantities, quickly and with high purity, resulting in high cost of separation and purification, long production time and other defects , In addition, the final biological products have more heterogeneous substances, and the side reactions are relatively large when used

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Separation and Purification of BHK21 Cells and Foot-and-Mouth Disease Virus Culture Mixture by Depth Filtration

[0043] Resuscitate BHK21 cells from the liquid nitrogen bank of cell working seeds, passage and expand in square bottles first, then expand and culture in turn bottle, 5L bioreactor, and 120L bioreactor in sequence, and then digest with trypsin to make cells suspension, inoculated in CLAVORUS TM Carry out large-scale expansion culture in 650L reactor (Beijing Tianherui Biotechnology Company), the working volume of bioreactor is 400L, cultivate after 5 days, inoculate foot-and-mouth disease virus on the BHK21 cell of gained, the cell in the bioreactor The culture medium was replaced with a virus culture medium, and the inoculated BHK21 cells were cultivated. After about 12-24 hours of cultivation in the bioreactor, the virus fluid was harvested at one time, and the harvested 400L culture mixture was stored in a liquid storage tank at 4°C at low te...

Embodiment 2

[0046] Example 2 Separation and Purification of Marc145 Cells and Porcine Reproductive and Respiratory Syndrome by Depth Filtration Virus (PRRS virus) culture mixture

[0047] Resuscitate Marc145 cells from the cell working seed liquid nitrogen bank, first passage and expand in the square bottle, then expand and culture in turn bottle, 5L bioreactor, and 120L bioreactor in sequence, and then digest with trypsin to make cells suspension, inoculated in CLAVORUS TM Carry out large-scale expansion culture in 650L reactor (Beijing Tianherui Biotechnology Company), the working volume of bioreactor is 300L, after cultivating 4 days, porcine reproductive and respiratory syndrome virus is inoculated on the obtained Marc145 cell, biological The cell culture medium in the reactor was replaced with the virus culture medium, and the inoculated Marc145 cells were cultured. After the virus was cultivated for 6 hours, the perfusion culture was started, and the virus liquid was harvested ...

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PUM

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Abstract

The invention discloses a separation and purification method for a cell culture mixture, comprising the step of: filtering the cell culture mixture by a filter, wherein the filter is provided with at least two layers of filtering membranes, the nominal pore diameter of each layer of filtering membrane is gradually reduced in the direction of flow of the filtrate, i.e. the nominal pore diameter of the first layer of filtering membrane is about 2-10mu m, and the nominal pore diameter of the final layer of filtering membrane is about 0.1-0.2mu m. The separation and purification method for a cell culture mixture disclosed by the invention has the advantages of strong pertinence, large treatment capacity, simple separation and purification step, stable product quality, low cost and the like.

Description

technical field [0001] The present invention relates to a method for separation and purification of biological products, in particular to a method for separation and purification of a cell culture mixture used to obtain target biological products. Background technique [0002] The separation and purification of the culture mixture of the target biological product is one of the most critical steps in the production of biological products. It is a pivotal step between harvesting the target culture mixture and further downstream processing, and has a direct impact on the yield, quality and reproducibility of biological products. It also determines the efficiency and cost of the overall production process of biological products. [0003] The separation and purification methods of the target culture mixture commonly used in the existing biological product production process are mostly the combined use of several methods such as centrifugation, gel chromatography, and filtration, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02C07K16/18C12R1/93
CPCC12N7/00C12N2710/00051C12N1/02
Inventor 陈文庆王建超刘俊生
Owner BEIJING SKYWING TECH CO LTD
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