Factorial designs combined with the steepest ascent method to optimize serum-free media for CHO cells

Chi Hsien Liu, I. Ming Chu*, Shiaw Min Hwang

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

35 Scopus citations

Abstract

A serum free medium for recombinant CHO NTHU 108 cell growth and fusion protein (CD20 linked to a human IgG-Fc γ4 fragment) synthesis were systematically developed using factorial designs combined with the steepest ascent method. Experimental results indicate that the optimal composition of serum replacement for specific fusion protein production was 1% SITE (selenium, insulin, transferrin, ethanolamine), 0.3 g/L yeast extract, and 0.09% linoleic acid-BSA. Cell growth and fusion protein production of the adapted CHO NTHU 108 cultured in Iscove's modified Dulbecco's medium supplemented with these serum substitutes were comparable to those in the Ex-Cell 301 commercial serum-free medium. These serum substitutes can also promote CHO cell growth and fusion protein production in nine kinds of commercial media. The low protein content of the developed medium facilitates downstream processing and product purification.

Original languageEnglish
Pages (from-to)314-321
Number of pages8
JournalEnzyme and Microbial Technology
Volume28
Issue number4-5
DOIs
StatePublished - 08 03 2001
Externally publishedYes

Keywords

  • CHO cells
  • Fusion protein
  • Serum substitutes
  • Serum-free media
  • Statistical design
  • Steepest ascent method

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