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Development of optimal medium for production of commercially important monoclonal antibody 520C9 by hybridoma cell

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Abstract

Hybridoma HB-8696 produces monoclonal antibody (mAb) 520C9 (mouse IgG1), which recognizes breast cancer oncoprotein c-erbB2. The objective of this study was to optimize the medium recipe of HB 8696 cell for production of mAb 520C9. The optimization consisted of two steps: (1) screening of significant nutrients to make subsequent experiments more efficient with less runs and (2) locating their optimal concentrations. 29 variables including essential and non-essential amino acids, glucose, serum and 6 salts, namely NaCl, KCl, CaCl2, NaH2PO4, MgSO4 and Na-pyruvate were chosen in screening phase. The Plackett–Burman method was used to screen the variables influencing mAb production. Seven factors namely glucose, serum, asparagine, threonine, serine, NaCl and NaH2PO4 were identified to have a positive influencing role on mAb production with a confidence level >90 % (p < 0.1). Finally, Response surface methodology revealed the optimal level of the variables. The mAb production and average specific mAb production rate were enhanced by 111.05 and 105 %, respectively, compared to control medium.

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Acknowledgments

We are thankful to Department of Biotechnology (DBT), Govt. of India, New Delhi for providing financial support during the work done.

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  1. Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology, Hauz Khas, New Delhi, 110016, India

    Sucharita Sen & Pradip K. Roychoudhury

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  1. Sucharita Sen
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  2. Pradip K. Roychoudhury
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Correspondence to Pradip K. Roychoudhury.

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Sen, S., Roychoudhury, P.K. Development of optimal medium for production of commercially important monoclonal antibody 520C9 by hybridoma cell. Cytotechnology 65, 233–252 (2013). https://doi.org/10.1007/s10616-012-9480-z

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