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Cell perfusion culture method for mini model based on Tubespin bioreactor

A technology of bioreactor and perfusion culture, which is applied in the field of cell perfusion culture of reduced models, can solve the problems of high cost and insufficient parallelism, and achieve the effects of low cost, easy operation and reduced shearing

Active Publication Date: 2019-08-09
HJB HANGZHOU CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current commercial perfusion shrinkage model is based on Amber15 TM The automated microreactor, which can achieve a maximum medium exchange volume of 2RV / day and a cell separation efficiency of more than 95%, is comparable to a large-scale perfusion reactor, but it has higher costs and parallel sexual insufficiency

Method used

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  • Cell perfusion culture method for mini model based on Tubespin bioreactor
  • Cell perfusion culture method for mini model based on Tubespin bioreactor
  • Cell perfusion culture method for mini model based on Tubespin bioreactor

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Experimental program
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Embodiment 1

[0049] The cell perfusion culture method based on the reduced model of Tubespin bioreactor of the present embodiment comprises the following steps:

[0050] (1) Expansion of seed cells: CHO cells stably expressing monoclonal antibody mAb are amplified in ActiPro containing 4mM Gln after recovery, passed once every three days, and placed in a 125-250mL conical shaker flask for culture and expansion. 37℃, rotation radius 25mm, speed 110rpm, humidity 80%, 5% CO 2 Infors shaker, when the cells grow to (3.0-4.0) × 10 6 cells / mL for next inoculation;

[0051] (2) The cells obtained in step (1) were divided into 0.5×10 6 cells / mL were inoculated into a 50mL Tubespin bioreactor, the volume of the cell liquid was 10mL, the angle of the Tubespin bioreactor was adjusted to 90° (the Tubespin bioreactor was perpendicular to the horizontal plane of the shaker), and no anti-shearing agent was added; Tubespin biological The reactor is cultivated at 37°C, the rotation radius is 50mm, the ro...

Embodiment 2

[0054] The cell perfusion culture method based on the reduced model of Tubespin bioreactor of the present embodiment comprises the following steps:

[0055] (1) Expansion of seed cells: CHO cells stably expressing monoclonal antibody mAb were amplified in ActiPro containing 4mMGln after recovery, passed once every three days, and placed in a 125-250mL Erlenmeyer shaker flask at 37 ℃, rotation radius 25mm, speed 110rpm, humidity 80%, 5% CO 2 Infors shaker, when the cells grow to (3.0-4.0) × 10 6 cells / mL for next inoculation;

[0056] (2) The cells obtained in step (1) were divided into 0.5×10 6 Inoculate the cells / mL into a 50mL Tubespin bioreactor, the cell liquid volume is 15mL, adjust the angle of the Tubespin bioreactor tube stand to 90° (the Tubespin bioreactor is perpendicular to the horizontal plane of the shaker), and do not add anti-shearing agent; Tubespin bioreactor culture at 37°C, rotation radius 50mm, rotation speed 190rpm, humidity 80%, 5% CO 2 Infors shaker...

Embodiment 3

[0059] The cell perfusion culture method based on the reduced model of Tubespin bioreactor of the present embodiment comprises the following steps:

[0060] (1) Expansion of seed cells: CHO cells stably expressing monoclonal antibody mAb are amplified in ActiPro containing 4mM Gln after recovery, passed once every three days, and placed in a 125-250mL conical shaker flask for culture and expansion. 37℃, rotation radius 25mm, speed 110rpm, humidity 80%, 5% CO 2 Infors shaker, when the cells grow to (3.0-4.0) × 10 6 cells / mL for next inoculation;

[0061] (2) The cells obtained in step (1) were divided into 0.5×10 6 cells / mL were inoculated into a 50mL Tubespin bioreactor, and the volume of the cell liquid was 10mL. Adjust the angle of the Tubespin bioreactor to 60° (60° between the Tubespin bioreactor and the horizontal plane of the shaker), without adding anti-shear reagent; Tubespin bioreactor culture at 37 ℃, rotation radius 50mm, rotation speed 190rpm, humidity 80%, 5% C...

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Abstract

The invention relates to the technical field of cell engineering of bioengineering, in particular to a cell perfusion culture method for a mini model based on a Tubespin bioreactor. The cell perfusionculture method for the mini model based on the Tubespin bioreactor comprises the following steps that cells are inoculated into the Tubespin bioreactor, the volume of cell sap accounts for 20-30% ofthe volume of the Tubespin bioreactor, the inclination angle of the Tubespin bioreactor is 60-90 degrees, and under the shaking condition, perfusion culture is conducted. According to the cell perfusion culture method, culture parameters of the Tubespin bioreactor are regulated and controlled, the ventilation amount of the Tubespin bioreactor is increased, shearing is reduced, and beneficial conditions are provided for high-density culture; meanwhile, the continuous perfusion culture process can be simulated, redundant cells are removed every day, centrifugation and liquid exchange are conducted, and the growth and yield of the cells are monitored, so that the culture process enters multiple steady states.

Description

technical field [0001] The invention relates to the technical field of cell engineering of bioengineering, in particular to a cell perfusion culture method based on a reduced model of a Tubespin bioreactor. Background technique [0002] Animal cell culture is a method that started in the early 20th century and is currently widely used in fields such as biology and medicine. Commonly used animal cell cultures are divided into batch culture, fed-batch culture, perfusion culture and so on. Among them, perfusion culture can continuously update the medium during the culture process, meet the nutritional needs of cells in time, and remove harmful metabolites such as lactic acid and ammonium ions. Therefore, this method effectively improves the animal cell density and viability, prolongs the culture time of the cells, and greatly increases the protein production. The significant advantage of perfusion culture is that the protein can be separated in time with the discharge of the ...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0682C12N2500/02C12N2527/00
Inventor 罗颖娣薛圣杰欧丽婷廖元霆
Owner HJB HANGZHOU CO LTD
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