Increased expression of the integral membrane proteins EGFR and FGFR3 in anti-apoptotic Chinese hamster ovary cell lines

Erika Ohsfeldt, Szu Han Huang, Deniz Baycin-Hizal, Linda Kristoffersen, Thuy My T. Le, Edwin Li, Kalina Hristova, Michael J. Betenbaugh

Research output: Contribution to journalArticlepeer-review

Abstract

Membrane proteins such as receptor tyrosine kinases (RTKs) have a vital role in many cellular functions, making them potential targets for therapeutic research. In this study, we investigated the coexpression of the anti-apoptosis gene Bcl-xL with model membrane proteins as a means of increasing membrane protein expression in mammalian cells. Chinese hamster ovary (CHO) cells expressing heterologous Bcl-xL and wild-type CHO cells were transfected with either epidermal growth factor receptor or fibroblast growth factor receptor 3. The CHO-Bcl-xL cell lines showed increased expression of both RTK proteins as compared with the wild-type CHO cell lines in transient expression analysis, as detected by Western blot and flow cytometry after 15 days of antibiotic selection in stable expression pools. Increased expression was also seen in clonal isolates from the CHO-Bcl-xL cell lines, whereas the clonal cell line expression was minimal in wild-type CHO cell lines. Our results demonstrate that application of the anti-apoptosis gene Bcl-xL can increase expression of RTK proteins in CHO cells. This approach may be applied to improve stable expression of other membrane proteins in the future using mammalian cell lines with Bcl-xL or perhaps other anti-apoptotic genes.

Original languageEnglish (US)
Pages (from-to)155-162
Number of pages8
JournalBiotechnology and Applied Biochemistry
Volume59
Issue number3
DOIs
StatePublished - May 1 2012

Keywords

  • Anti-apoptosis
  • Bcl-2 protein family
  • Growth factor receptors
  • Mammalian cell culture
  • Membrane protein expression
  • Tyrosine kinases

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Molecular Medicine
  • Biomedical Engineering
  • Applied Microbiology and Biotechnology
  • Drug Discovery
  • Process Chemistry and Technology

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