Enhancement of recombinant protein production in Chinese hamster ovary cells through anti-apoptosis engineering using 30Kc6 gene

Shin Sik Choi, Won Jong Rhee, Eun Jeong Kim, Tai Hyun Park

Research output: Contribution to journalArticlepeer-review

60 Scopus citations

Abstract

It was previously reported that silkworm hemolymph (SH) inhibits apoptosis and increases the production of recombinant human erythropoietin (EPO) in Chinese hamster ovary (CHO) cells. The apoptosis-inhibiting component in SH is a member of 30K protein family. In this study, the CHO cell line producing EPO was manipulated genetically to express the 30Kc6 gene encoding a 30K protein in the hemolymph of the silkworm, Bombyx mori. The transient expression of 30Kc6 significantly suppressed the cell death induced by serum deprivation. A stable cell line expressing 30Kc6 with an anti-apoptotic property was established. The stable expression of 30Kc6 inhibited serum-deprivation-induced apoptosis and increased the cell density and EPO titer by 5- and 10-fold, respectively. The positive effects of the 30Kc6 expression on cell viability and productivity were due to the stable maintenance of the mitochondrial activity. The 30Kc6 expression efficiently suppressed the depolarization of the mitochondrial membrane and subsequently balanced the generation/consumption of ATP. The use of the 30Kc6 gene is expected to provide a new method of host cell engineering for improving the productivity of the recombinant protein.

Original languageEnglish
Pages (from-to)459-467
Number of pages9
JournalBiotechnology and Bioengineering
Volume95
Issue number3
DOIs
StatePublished - 20 Oct 2006

Keywords

  • 30Kc6
  • Apoptosis
  • CHO
  • EPO
  • Recombinant protein production
  • Silkworm

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