ABSTRACT Low cost and high efficiency make disposable bioreactors feasible for small-scale therapeutic development and initial clinical trials. We have developed a cryogel-based disposable

bioreactor matrix, which has been used for production of protein therapeutics such as urokinase and monoclonal antibodies (mAbs). The protocol discusses the application of a cryogel

bioreactor for mAb production. Cryogels composed of either polyacrylamide (PAAm) coupled to gelatin or semi-interpenetrating PAAm-chitosan are synthesized by free-radical polymerization at

−12 °C. Hybridoma cells are immobilized over the cryogel bioreactor and incubated for 48 h. Medium is circulated thereafter at 0.2 ml min−1 and bioreactors can be run continuously for 60 d.

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42, 895–900 (1979). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We acknowledge the financial support from the Department of Biotechnology, Ministry of Science and

Technology, Government of India; Protista Biotechnology Sweden; and the Swedish Research Council/SIDA Research Link Project. We are also thankful to all other co-workers, especially V.

Bansal, S. Nilsang and B. Mattiasson who have contributed to the development of this work in one way or another. We also thank A. Karande for her kind help in providing the hybridoma cell

lines and for mAb analysis. AUTHOR INFORMATION Author notes * Era Jain Present address: Present address: Department of Chemical Engineering, Virginia Polytechnic Institute and State

University, Blacksburg, Virginia, USA., AUTHORS AND AFFILIATIONS * Department of Biological Sciences and Bioengineering, Indian Institute of Technology Kanpur, Kanpur, India Era Jain & 

Ashok Kumar Authors * Era Jain View author publications You can also search for this author inPubMed Google Scholar * Ashok Kumar View author publications You can also search for this author

inPubMed Google Scholar CONTRIBUTIONS Both the authors (E.J. and A.K.) conceived, planned and conducted experiments on different aspects of this study. A.K. was responsible for supervising

all the work described in this protocol. CORRESPONDING AUTHOR Correspondence to Ashok Kumar. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests.

SUPPLEMENTARY INFORMATION SUPPLEMENTARY METHODS Determination of Monoclonal Antibody; Analysis of Glucose in spent media samples; Determination of Ammonia; Simultaneous Determination of

Glucose, Lactic acid, and Glutamine by HPLC; and Determination of Urokinase Activity (PDF 369 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Jain,

E., Kumar, A. Disposable polymeric cryogel bioreactor matrix for therapeutic protein production. _Nat Protoc_ 8, 821–835 (2013). https://doi.org/10.1038/nprot.2013.027 Download citation *

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