Overexpression of heat shock proteins (HSPs) in CHO cells for extended culture viability and improved recombinant protein production

Lee, Yih Yean; Wong, Kainam Thomas; Tan, Jiafu; Toh, Poh Choo; Mao, Yanying; Brusic, Vesna; Yap, Melvin J.

doi:10.1016/j.jbiotec.2009.05.013

Cited by 44 publications

(38 citation statements)

References 29 publications

(27 reference statements)

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“…Among many factors affecting mAbs quality, the stability with time of high producing level CHO cell clones with enhanced endogenous pathways (e.g glutamine synthetase (GS) gene) [3], and presenting a prolonged cell viability level due to the over-expression of some cytoplasmic proteins (e.g. chaperones such as Hsp70 and Hsp27) is highly critical [4]. Moreover, media composition and culture conditions, as well as their management along with culture duration, have to be optimized to achieve not only the objective of desired cell productivity and viability but also mAbs quality specifications [5].…”

Section: Introductionmentioning

confidence: 99%

Analyzing Clonal Variation of Monoclonal Antibody-Producing CHO Cell Lines Using an In Silico Metabolomic Platform

et al. 2014

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Monoclonal antibody producing Chinese hamster ovary (CHO) cells have been shown to undergo metabolic changes when engineered to produce high titers of recombinant proteins. In this work, we have studied the distinct metabolism of CHO cell clones harboring an efficient inducible expression system, based on the cumate gene switch, and displaying different expression levels, high and low productivities, compared to that of the parental cells from which they were derived. A kinetic model for CHO cell metabolism was further developed to include metabolic regulation. Model calibration was performed using intracellular and extracellular metabolite profiles obtained from shake flask batch cultures. Model simulations of intracellular fluxes and ratios known as biomarkers revealed significant changes correlated with clonal variation but not to the recombinant protein expression level. Metabolic flux distribution mostly differs in the reactions involving pyruvate metabolism, with an increased net flux of pyruvate into the tricarboxylic acid (TCA) cycle in the high-producer clone, either being induced or non-induced with cumate. More specifically, CHO cell metabolism in this clone was characterized by an efficient utilization of glucose and a high pyruvate dehydrogenase flux. Moreover, the high-producer clone shows a high rate of anaplerosis from pyruvate to oxaloacetate, through pyruvate carboxylase and from glutamate to α-ketoglutarate, through glutamate dehydrogenase, and a reduced rate of cataplerosis from malate to pyruvate, through malic enzyme. Indeed, the increase of flux through pyruvate carboxylase was not driven by an increased anabolic demand. It is in fact linked to an increase of the TCA cycle global flux, which allows better regulation of higher redox and more efficient metabolic states. To the best of our knowledge, this is the first time a dynamic in silico platform is proposed to analyze and compare the metabolomic behavior of different CHO clones.

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Section: Introductionmentioning

confidence: 99%

Analyzing Clonal Variation of Monoclonal Antibody-Producing CHO Cell Lines Using an In Silico Metabolomic Platform

et al. 2014

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“…Optimizing culture conditions are important for the expression system [22,23], such as cell density [24], cell viability [25], the glucose concentration [26][27][28]. Excessive or insufficient nutrients can influence cell growth, which finally leads to low production of interesting proteins.…”

Section: Discussionmentioning

confidence: 99%

Efficient production of FAM19A4, a novel potential cytokine, in a stable optimized CHO-S cell system

Liu

Wang

Shi

et al. 2015

Protein Expression and Purification

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“…Heat shock proteins (HSPs) belong to a class of highly conserved proteins that act physiologically as molecular chaperones to stabilize existing proteins against aggregation and mediate the folding of newly translated proteins in the cytosol and other organelles [23]. Additionally, they have been shown to demonstrate antiapoptotic effects against a wide range of both physical and chemical apoptotic stimuli [24].…”

Section: Discussionmentioning

confidence: 99%

Gamma-Tocotrienol Modulated Gene Expression in Senescent Human Diploid Fibroblasts as Revealed by Microarray Analysis

Makpol

Zainuddin

Chua

et al. 2013

Oxidative Medicine and Cellular Longevity

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The effect of γ-tocotrienol, a vitamin E isomer, in modulating gene expression in cellular aging of human diploid fibroblasts was studied. Senescent cells at passage 30 were incubated with 70 μM of γ-tocotrienol for 24 h. Gene expression patterns were evaluated using Sentrix HumanRef-8 Expression BeadChip from Illumina, analysed using GeneSpring GX10 software, and validated using quantitative RT-PCR. A total of 100 genes were differentially expressed (P < 0.001) by at least 1.5 fold in response to γ-tocotrienol treatment. Amongst the genes were IRAK3, SelS, HSPA5, HERPUD1, DNAJB9, SEPR1, C18orf55, ARF4, RINT1, NXT1, CADPS2, COG6, and GLRX5. Significant gene list was further analysed by Gene Set Enrichment Analysis (GSEA), and the Normalized Enrichment Score (NES) showed that biological processes such as inflammation, protein transport, apoptosis, and cell redox homeostasis were modulated in senescent fibroblasts treated with γ-tocotrienol. These findings revealed that γ-tocotrienol may prevent cellular aging of human diploid fibroblasts by modulating gene expression.

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Overexpression of heat shock proteins (HSPs) in CHO cells for extended culture viability and improved recombinant protein production

Cited by 44 publications

References 29 publications

Analyzing Clonal Variation of Monoclonal Antibody-Producing CHO Cell Lines Using an In Silico Metabolomic Platform

Analyzing Clonal Variation of Monoclonal Antibody-Producing CHO Cell Lines Using an In Silico Metabolomic Platform

Efficient production of FAM19A4, a novel potential cytokine, in a stable optimized CHO-S cell system

Gamma-Tocotrienol Modulated Gene Expression in Senescent Human Diploid Fibroblasts as Revealed by Microarray Analysis

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