Efficient expression and isolation of recombinant human interleukin-11 (rhIL-11) in Pichia pastoris

Kuo Ming Yu, Johnson Yiu-Nam Lau, Manson Fok, Yuk Keung Yeung, Siu Ping Fok, Felix Shek, Wing Tak Wong, Qui Lim Choo

Research output: Journal article publicationJournal articleAcademic researchpeer-review

10 Citations (Scopus)

Abstract

Current source of recombinant human interleukin-11 (rhIL-11) is isolated from a fusion protein expressed by E. coli that requires additional enterokinase to remove linked protein, resulting in product heterogeneity of N-terminal sequence. Due to lack of glycosylation, rhIL-11 is suitable to be expressed by yeast cells. However, the only available yeast-derived rhIL-11 presents an obstacle in low production yield, as well as an unamiable process, such as the use of reverse-phase chromatography employing plenty of toxic organic solvents. Our findings showed that the low yield was due to self-aggregation of rhIL-11. A novel process recovering bioactive rhIL-11 from the yeast secretory medium therefore has been developed and demonstrated, involving fermentation from Pichia pastoris, followed by a two-phase extraction to precipitate rhIL-11. After renaturing, the protein of interest was purified by a two-column step, comprising a cation-exchanger, and a hydrophobic interaction chromatography in tandem at high sample loads that was facile and cost-effective in future scale-up. Identity and quality assessments confirmed the expected amino acid sequence without N-terminal heterogeneity, as well as high quality in potency and purity. Such a process provides an alternative and adequate supply of the starting material for the PEGylated rhIL-11.

Original languageEnglish
Pages (from-to)69-77
Number of pages9
JournalProtein Expression and Purification
Volume146
DOIs
Publication statusPublished - Jun 2018

Keywords

  • IL-11
  • PEG
  • Secretory
  • Soluble aggregate
  • Yeast

ASJC Scopus subject areas

  • Biotechnology

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