Rapid protein production using CHO stable transfection pools

Biotechnology Progress
Jianxin YeJulia F Markusen

Abstract

During early preclinical development of therapeutic proteins, representative materials are often required for process development, such as for pharmacokinetic/pharmacodynamic studies in animals, formulation design, and analytical assay development. To rapidly generate large amounts of representative materials, transient transfection is commonly used. Because of the typical low yields with transient transfection, especially in CHO cells, here we describe an alternative strategy using stable transfection pool technology. Using stable transfection pools, gram quantities of monoclonal antibody (Mab) can be generated within 2 months post-transfection. Expression levels for monoclonal antibodies can be achieved ranging from 100 mg/L to over 1000 mg/L. This methodology was successfully scaled up to a 200 L scale using disposable bioreactor technology for ease of rapid implementation. When fluorescence-activated cell sorting was implemented to enrich the transfection pools for high producers, the productivity could be improved by about three-fold. We also found that an optimal production time window exists to achieve the highest yield because the transfection pools were not stable and productivity generally decreased over length in cul...Continue Reading

References

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Citations

Dec 10, 2013·FEBS Letters·Ashok D Bandaranayake, Steven C Almo
Oct 5, 2011·Biotechnology Advances·Jianwei Zhu
May 16, 2015·Nucleic Acids Research·Katalin ZborayEmilio Casanova
Feb 13, 2014·New Biotechnology·Jeff Jia Cheng HouTrent P Munro
Mar 12, 2015·Journal of Biotechnology·Sowmya BalasubramanianFlorian M Wurm
Dec 1, 2015·Biotechnology and Bioengineering·Sowmya BalasubramanianFlorian M Wurm
Jun 9, 2016·Biotechnology Journal·Joseph J PriolaMichael J Betenbaugh
Sep 30, 2016·Journal of Immunological Methods·Jared L SpidelJ Bradford Kline
Jun 20, 2017·Journal of Biotechnology·Adeline PoulainYves Durocher
Dec 15, 2017·Nature Protocols·Rodrigo Vazquez-LombardiDaniel Christ
May 7, 2020·Applied Microbiology and Biotechnology·Tian-Yun Wang, Xiao Guo
Sep 24, 2019·Antibodies·Qiong WangMichael J Betenbaugh

Related Concepts

Monoclonal Antibodies
Cricetulus
Flow Cytometry
Protein Glycosylation
Cricetus
Transfection
Protein Engineering
Chinese Hamster Ovary Cell
Monoclonal Antibodies
Disease Vectors

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