Improved process for production of recombinant yeast-derived monomeric human G-CSF

C. S. Bae, D. S. Yang, Jeewon Lee, Y. H. Park

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

The human granulocyte colony-stimulating factor (hG-CSF) was efficiently secreted at high levels in fed-batch cultures of recombinant Saccharomyces cerevisiae. However, the secreted recombinant hG-CSF (rhG-CSF) was shown to exist as large multimers in the culture broth due to strong hydrophobic interaction. It was hardly monomerized even by urea at high concentration. This multimer has been reported to diminish specific receptor-binding activity of hG-CSF and causes undesirable problems in the downstream process. When the rhG-CSF was secreted to extracellular broth in the presence of a non-ionic surfactant (Tween 80) in the culture media, the multimerization of the secreted rhGCSF was efficiently prevented in the fed-batch cultures. Also, the monomer fraction and secretion efficiency of rhG-CSF were significantly increased at the higher culture pH (6.5). Without using any denaturing agents, the secreted rhG-CSF monomer was easily purified with high recovery yield and purity via a simple purification process under acidic conditions, consisting of diafiltration, cation exchange, and gel filtration chromatography. A lyophilization process devoid of intermonomer aggregation was also designed using effective stabilizing agents.

Original languageEnglish
Pages (from-to)338-344
Number of pages7
JournalApplied Microbiology and Biotechnology
Volume52
Issue number3
DOIs
Publication statusPublished - 1999 Sep
Externally publishedYes

Fingerprint

Granulocyte Colony-Stimulating Factor
Yeast
Batch Cell Culture Techniques
Yeasts
Monomers
Nonionic surfactants
Chromatography
Urea
Purification
Ion exchange
Freeze Drying
Gels
Agglomeration
Polysorbates
Excipients
Positive ions
Hydrophobic and Hydrophilic Interactions
Human Activities
Surface-Active Agents
Recovery

ASJC Scopus subject areas

  • Biotechnology
  • Microbiology
  • Bioengineering

Cite this

Improved process for production of recombinant yeast-derived monomeric human G-CSF. / Bae, C. S.; Yang, D. S.; Lee, Jeewon; Park, Y. H.

In: Applied Microbiology and Biotechnology, Vol. 52, No. 3, 09.1999, p. 338-344.

Research output: Contribution to journalArticle

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