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Cytotechnology

, Volume 63, Issue 3, pp 247–258 | Cite as

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

  • Suwen Hu
  • Lei Deng
  • Huamao Wang
  • Yingping ZhuangEmail author
  • Ju Chu
  • Siliang Zhang
  • Zhonghai Li
  • Meijin GuoEmail author
Original Research

Abstract

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 1 mM, 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 44.72 mg/g and the mean yield coefficient of glutamine to antibody was 721.40 mg/g. Finally, in small-scale bioreactor culture, the highest total amount of C12 antibody (1,854 mg) 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.

Keywords

Mouse-human chimeric anti-EGFRvIII antibody Recombinant CHO-C12 cells Balanced amino acid feed Perfusion culture 

Notes

Acknowledgments

This study was supported by the National High Technology Research & Development Program (863 Program) of China (No. 2007AA02Z216), the National Special Fund for State Key Laboratory of Bioreactor Engineering (No.2060204),the National Basic Research Program of China (No.2007CB714303) and the Qianjiang Scholarship grant from Hangzhou Municipal Government, China.

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Suwen Hu
    • 1
  • Lei Deng
    • 1
  • Huamao Wang
    • 2
  • Yingping Zhuang
    • 1
    Email author
  • Ju Chu
    • 1
  • Siliang Zhang
    • 1
  • Zhonghai Li
    • 2
  • Meijin Guo
    • 1
    Email author
  1. 1.State Key Laboratory of Bioreactor EngineeringEast China University of Science and TechnologyShanghaiPeople’s Republic of China
  2. 2.State Key Laboratory of Oncogenes and Related GenesShanghai Jiaotong UniversityShanghaiPeople’s Republic of China

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