Efficient production process of bioactive recombinant human leukemia inhibitory factor in Chinese hamster ovary cells

Sujin Cho, Gookjoo Jeong, Nara Han, Changin Kim, Jeong Soo Park, Yongsu Jeong, Kwanghee Baek, Jaeseung Yoon

Research output: Contribution to journalArticlepeer-review

Abstract

The rhLIF is widely used as an essential factor in stem cell cultures for cell therapies. However, all the recombinant LIFs commercially available are expensive, and no commercially available rhLIF meet the standards recommended by USP for use in cell therapies. The current study reports the efficient production of N-glycosylated and bioactive rhLIF in CHO cells. The production rate of established rhLIF-expressing rCHO cells was approximately 0.85 g/l in 12-day fed-batch cultures using a 7.5 l bioreactor. The rhLIF protein was purified via a four-step purification procedure with approximately 57% recovery rate and greater than 99% purity. The purified rhLIF was N-glycosylated and biologically active with an EC50 of 0.167 ng/ml and a specific activity of 0.86 × 103 IU/mg. The purification procedure controlled the levels of process-related impurities below critical levels recommended by USP for cytokines used in cell therapies. The current work is the first production process of N-glycosylated and bioactive rhLIF, which can be applied to large-scale manufacture of GMP-grade rhLIF for use as an ancillary material in cell therapy.

Original languageEnglish
Article number105744
JournalProtein Expression and Purification
Volume176
DOIs
Publication statusPublished - Dec 2020

Keywords

  • Bioassay
  • Bioreactor process
  • Chinese hamster ovary (CHO) cells
  • Leukemia inhibitory factor (LIF)
  • Purification
  • Recombinant protein expression
  • USP

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