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Methods for improving cell viability in production bioreactor

A bioreactor, cell technology, applied in the fields of biotechnology and cell biology, can solve the problems of cell shear sensitivity and increased cell death

Pending Publication Date: 2020-12-25
GENZYME CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the addition and accumulation of antifoams may make cells shear-sensitive, leading to increased cell death

Method used

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  • Methods for improving cell viability in production bioreactor
  • Methods for improving cell viability in production bioreactor
  • Methods for improving cell viability in production bioreactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0273] Example 1: Combination of full factorial screening of test culture vessels with an endocentric composite design

[0274] Select rotary agitation (RPM 3 ), sensitizer concentration and protectant concentration as parameters. A full factorial design combined with an endocentral composite design was used to determine a set of test conditions ( Figure 7). Specific LDH production values ​​were measured for each test condition. The results are used to provide estimated contours for the selected parameters. Identify areas of poor performance and operational failures ( Figure 9 ).

Embodiment 2

[0275] Example 2: Prediction of Specific LDH Production Values ​​at Bench Scale

[0276] A proof-of-concept experiment was performed at bench scale to determine the accuracy of the prediction method as described herein.

[0277] The protectant concentration in bench scale was set at three different levels. For each level, two benchtop culture bioreactors were tested. Measure the peak value of specific LDH production due to the perfusion process and compare it to the predicted value.

[0278] Actual observations from bench-scale experiments matched those predicted very well (P2 =0.84)( Figure 13A and Figure 13C ). It turns out that the method as described here can be successfully used to predict the impact of multiple parameters at higher scales.

[0279] Experiments were conducted to maintain the defoamer concentration in the range of 30-40 ppm defoamer addition per day. Pluronic concentration was adjusted from 1.8g / L (dotted line), 2.8g / L (solid line, filled circle), ...

Embodiment 3

[0281] Embodiment 3: Predict the specific LDH output value under the production scale

[0282] Experiments were performed at production scale to determine the accuracy of the models as described herein.

[0283] Three 50L production bioreactors and two 100L production bioreactors were tested. Poloxamer-188 (Pluronic) concentrations ranging from 1.8 g / L to 5.8 g / L were experimentally tested.

[0284] The actual observed values ​​for specific LDH production matched these predicted values ​​very well (P2 =0.84)( Figure 15A and Figure 15B ). The results demonstrate that the method as described herein can be successfully used to predict the impact of selected parameters at production scale (production bioreactor scale).

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Abstract

Provided herein are methods of predicting the effect of a concentration of a sensitizer on cell viability in a production bioreactor, methods of improving cell viability in a production bioreactor, methods of predicting cell viability in a production bioreactor, and methods for culturing a cell in a production bioreactor.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 644,339, filed March 16, 2018, and U.S. Provisional Patent Application Serial No. 62 / 645,755, filed March 20, 2018; The entire contents are incorporated herein by reference. technical field [0003] This disclosure relates to methods of increasing cell viability in production bioreactors and to the fields of biotechnology and cell biology. Background technique [0004] Process intensification offers many advantages for the production of recombinant therapeutic proteins. One of the strategies for process intensification is to maintain high cell density to improve recombinant protein production, which also requires high oxygen demand. Oxygen sparging in the production bioreactor results in foaming in the production bioreactor. Foaming typically occurs during fortified fed-batch and perfusion. Therefore, defoamers are often added dur...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50C12M1/34
CPCG01N33/5014C12M41/46C12Q1/32C12Q1/025C12M41/32C12M27/20C12M47/12C12M41/42
Inventor J·王N·沙J·沃尔特陆久怡T·约翰逊任玉坤J·麦卡蒂
Owner GENZYME CORP
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