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Hybridoma HB-8696 produces monoclonal antibody (mAb) 520C9 (mouse IgG(1)), 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, CaCl(2), NaH(2)PO(4), MgSO(4) 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 NaH(2)PO(4) were identified to have a positive influencing role on mAb production with a confidence level &gt;90 % (p &lt; 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.

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