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

被引：18

作者：

Lee, Kyoung Ho ^{[1
]}

Tsutsui, Tomomi ^{[2
]}

Honda, Kohsuke ^{[1
]}

Asano, Ryutaro ^{[3
]}

Kumagai, Izumi ^{[3
]}

Ohtake, Hisao ^{[1
]}

Omasa, Takeshi ^{[1
,2
]}

机构：

[1] Osaka Univ, Grad Sch Engn, Dept Biotechnol, Suita, Osaka 5650871, Japan

[2] Univ Tokushima, Inst Sci & Technol, Tokushima 7708506, Japan

[3] Tohoku Univ, Grad Sch Engn, Dept Biomol Engn, Aoba Ku, Sendai, Miyagi 9808579, Japan

来源：

JOURNAL OF BIOSCIENCE AND BIOENGINEERING | 2013年 / 116卷 / 06期

基金：

日本学术振兴会;

关键词：

Chinese hamster ovary cells; Gene amplification; Monoclonal antibody; CDC25A; Cell cycle; DIHYDROFOLATE-REDUCTASE GENE; CDC25A PHOSPHATASE; CHO-CELLS; CHROMOSOME REARRANGEMENT; G(1)/S TRANSITION; MAMMALIAN-CELLS; AMPLIFICATION; EXPRESSION; ANTIBODY; PHOSPHORYLATION;

D O I：

10.1016/j.jbiosc.2013.05.032

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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 250 nM methotrexate (MTX). Cell cycle analysis revealed that the G2 to M phase transition rate was increased similar to 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. (C) 2013, The Society for Biotechnology, Japan. All rights reserved.

引用

页码：754 / 760

页数：7

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