Generation of high-producing cell lines by overexpression of cell division cycle 25 homolog A in Chinese hamster ovary cells

Kyoung Ho Lee, Tomomi Tsutsui, Kohsuke Honda, Ryutaro Asano, Izumi Kumagai, Hisao Ohtake, Takeshi Omasa

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

To improve the efficiency of conventional gene amplification systems, the effect of cell cycle modification during the gene amplification process on IgG production was investigated in Chinese hamster ovary (CHO) cells. The full-length cDNA of CHO cell division cycle 25 homolog A (Cdc25A) was introduced into CHO DG44 cells and the effects of CDC25A overexpression on the cell cycle, transgene copy number and IgG productivity were examined. Both wild-type and mutated CDC25A-overexpressing CHO cells showed a rapid increase in transgene copy number compared with mock cells during the gene amplification process, in both cell pools and individual clones. High-producing clones were obtained with high frequency in CDC25A-overexpressing cell pools. The specific production rate of the isolated clone CHO SD-S23 was up to 2.9-fold higher than that of mock cells in the presence of 250nM methotrexate (MTX). Cell cycle analysis revealed that the G2 to M phase transition rate was increased ~1.5-fold in CDC25A-overexpressing CHO cells under MTX treatment. Our results show the improvement of conventional gene amplification systems via cell cycle engineering at an early stage of cell line development.

Original languageEnglish
Pages (from-to)754-760
Number of pages7
JournalJournal of Bioscience and Bioengineering
Volume116
Issue number6
DOIs
Publication statusPublished - 2013 Dec

Keywords

  • CDC25A
  • Cell cycle
  • Chinese hamster ovary cells
  • Gene amplification
  • Monoclonal antibody

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint

Dive into the research topics of 'Generation of high-producing cell lines by overexpression of cell division cycle 25 homolog A in Chinese hamster ovary cells'. Together they form a unique fingerprint.

Cite this