Improving expression of recombinant human IGF-1 using IGF-1R knockout CHO cell lines

被引：11

作者：

Romand, Sandrine ^{[1
]}

Jostock, Thomas ^{[1
]}

Fornaro, Mara ^{[2
]}

Schmidt, Joerg ^{[3
]}

Ritter, Anett ^{[1
]}

Wilms, Burkhard ^{[1
]}

Laux, Holger ^{[1
]}

机构：

[1] Novartis Pharmaceut, Integrated Biol Profiling, Klybeckstr 141, CH-4057 Basel, Switzerland

[2] Novartis Inst BioMed Res, Musculoskeletal Dis, Basel, Switzerland

[3] Novartis Pharmaceut, Cell & Gene Therapies, Basel, Switzerland

来源：

BIOTECHNOLOGY AND BIOENGINEERING | 2016年 / 113卷 / 05期

关键词：

CHO; IGF-1; IGF-1R; cell line engineering; ZFN; Chinese hamster ovary; GROWTH-FACTOR-I; MONOCLONAL-ANTIBODY PRODUCTION; SOMATIC MAMMALIAN CELLS; HAMSTER OVARY CELLS; PROTEIN THERAPEUTICS; INSULIN; RECEPTOR; APOPTOSIS; GENES; CULTIVATION;

D O I：

10.1002/bit.25877

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：

Chinese Hamster Ovary (CHO) cells are widely used for the large-scale production of recombinant biopharmaceuticals. However, attempts to express IGF-1 (a mutated human Insulin-like growth factor 1 Ea peptide (hIGF-1Ea mut)) in CHO cells resulted in poor cell growth and low productivity (0.1-0.2g/L). Human IGF-1 variants negatively impacted CHO cell growth via the IGF-1 receptor (IGF-1R). Therefore knockout (KO) of the IGF-1R gene in two different CHO cell lines as well as knockdown (KD) of IGF-1R in one CHO cell line were performed. These cell line engineering approaches decreased significantly the hIGF-1 mediated cell growth inhibition and increased productivity of both KO CHO cell lines as well as of the KD CHO cell line. A productivity increase of 10-fold at pool level and sevenfold at clone level was achieved, resulting in a titer of 1.3g/L. This data illustrate that cell line engineering approaches are powerful tools to improve the yields of recombinant proteins which are difficult to produce in CHO cells. Biotechnol. Bioeng. 2016;113: 1094-1101. (c) 2015 Wiley Periodicals, Inc.

引用

页码：1094 / 1101

页数：8

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