caa a22 4500 44582316X CHVBK 20180317145250.0 cr unu---uuuuu 170323e20110501xx s 000 0 eng 10.1007/s10616-011-9336-y doi (NATIONALLICENCE)springer-10.1007/s10616-011-9336-y Bioprocess development for the production of mouse-human chimeric anti-epidermal growth factor receptor vIII antibody C12 by suspension culture of recombinant Chinese hamster ovary cells [Elektronische Daten] [Suwen Hu, Lei Deng, Huamao Wang, Yingping Zhuang, Ju Chu, Siliang Zhang, Zhonghai Li, Meijin Guo] The mouse-human chimeric anti-epidermal growth factor receptor vIII (EGFRvIII) antibody C12 is a promising candidate for the diagnosis of hepatocellular carcinoma (HCC). In this study, 3 processes were successfully developed to produce C12 by cultivation of recombinant Chinese hamster ovary (CHO-DG44) cells in serum-free medium. The effect of inoculum density was evaluated in batch cultures of shaker flasks to obtain the optimal inoculum density of 5×105 cells/mL. Then, the basic metabolic characteristics of CHO-C12 cells were studied in stirred bioreactor batch cultures. The results showed that the limiting concentrations of glucose and glutamine were 6 and 1mM, respectively. The culture process consumed significant amounts of aspartate, glutamate, asparagine, serine, isoleucine, leucine, and lysine. Aspartate, glutamate, asparagine, and serine were particularly exhausted in the early growth stage, thus limiting cell growth and antibody synthesis. Based on these findings, fed-batch and perfusion processes in the bioreactor were successfully developed with a balanced amino acid feed strategy. Fed-batch and especially perfusion culture effectively maintained high cell viability to prolong the culture process. Furthermore, perfusion cultures maximized the efficiency of nutrient utilization; the mean yield coefficient of antibody to consumed glucose was 44.72mg/g and the mean yield coefficient of glutamine to antibody was 721.40mg/g. Finally, in small-scale bioreactor culture, the highest total amount of C12 antibody (1,854mg) was realized in perfusion cultures. Therefore, perfusion culture appears to be the optimal process for small-scale production of C12 antibody by rCHO-C12 cells. Springer Science+Business Media B.V., 2011 Mouse-human chimeric anti-EGFRvIII antibody nationallicence Recombinant CHO-C12 cells nationallicence Balanced amino acid feed nationallicence Perfusion culture nationallicence Hu Suwen State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut Deng Lei State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut Wang Huamao State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiaotong University, 200032, Shanghai, People's Republic of China aut Zhuang Yingping State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut Chu Ju State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut Zhang Siliang State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut Li Zhonghai State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiaotong University, 200032, Shanghai, People's Republic of China aut Guo Meijin State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut Cytotechnology Springer Netherlands 63/3(2011-05-01), 247-258 0920-9069 63:3<247 2011 63 10616 https://doi.org/10.1007/s10616-011-9336-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10616-011-9336-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hu Suwen State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Deng Lei State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Wang Huamao State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiaotong University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Zhuang Yingping State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Chu Ju State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Siliang State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Li Zhonghai State Key Laboratory of Oncogenes and Related Genes, Shanghai Jiaotong University, 200032, Shanghai, People's Republic of China aut NATIONALLICENCE 700 1- Guo Meijin State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Rd., P.O. box 329, 200237, Shanghai, People's Republic of China aut NATIONALLICENCE 773 0- Cytotechnology Springer Netherlands 63/3(2011-05-01), 247-258 0920-9069 63:3<247 2011 63 10616 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer