Cytotechnology

, Volume 65, Issue 3, pp 363–378

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

  • Huifeng Zhang
  • Haibin Wang
  • Mei Liu
  • Tao Zhang
  • Ji Zhang
  • Xiangjing Wang
  • Wensheng Xiang
Original Research

DOI: 10.1007/s10616-012-9488-4

Cite this article as:
Zhang, H., Wang, H., Liu, M. et al. Cytotechnology (2013) 65: 363. doi:10.1007/s10616-012-9488-4

Abstract

A serum-free medium (CHO-SFM) together with a fed-batch process was developed for the cultivation of a recombinant GS-CHO cell line producing TNFR-Fc. According to the metabolic characteristics of GS-CHO cell, a basal medium was prepared by supplementing DMEM:F12:RPMI1640 (2:1:1) with amino acids, insulin, transferrin, Pluronic F68 and some other ingredients. Statistical optimization approaches based on Plackett–Burman and central composite designs were then adopted to identify additional positive determinants and determine their optimal concentrations, which resulted in the final CHO-SFM medium formulations. The maximum antibody titer reached was 90.95 mg/l in the developed CHO-SFM, which was a 18 % and 10 fold higher than that observed in the commercial EX-CELL™ 302 medium (76.95 mg/l) and basal medium (8.28 mg/l), respectively. Subsequently, a reliable, reproducible and robust fed-batch strategy was designed according to the offline measurement of glucose, giving a final antibody yield of 378 mg/l, which was a threefold improvement over that in conventional batch culture (122 mg/l) using CHO-SFM. In conclusion, the use of design of experiment (DoE) method facilitated the development of CHO-SFM medium and fed-batch process for the production of recombinant antibody using GS-CHO cells.

Keywords

GS-CHO cell Recombinant antibody SFM Design of experiment (DoE) Fed-batch process 

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Huifeng Zhang
    • 1
  • Haibin Wang
    • 2
  • Mei Liu
    • 1
  • Tao Zhang
    • 1
  • Ji Zhang
    • 1
  • Xiangjing Wang
    • 1
  • Wensheng Xiang
    • 1
  1. 1.School of Life ScienceNortheast Agricultural UniversityHarbinChina
  2. 2.Zhejiang Hisun Pharmaceutical Co., Ltd.TaizhouChina

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