scholarly journals Optimization of gene delivery methods in Xenopus laevis kidney (A6) and Chinese hamster ovary (CHO) cell lines for heterologous expression of Xenopus inner ear genes

2011 ◽  
Vol 47 (9) ◽  
pp. 640-652 ◽  
Author(s):  
Daniel Ramirez-Gordillo ◽  
Casilda Trujillo-Provencio ◽  
V. Bleu Knight ◽  
Elba E. Serrano
Keyword(s):  
Gene Delivery ◽  
Xenopus Laevis ◽  
Heterologous Expression ◽  
Inner Ear ◽  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Cho Cell ◽  
Delivery Methods

scholarly journals Sustained productivity in recombinant Chinese Hamster Ovary (CHO) cell lines: proteome analysis of the molecular basis for a process-related phenotype

2011 ◽  
Vol 11 (1) ◽  
Author(s):  
Paula Meleady ◽  
Padraig Doolan ◽  
Michael Henry ◽  
Niall Barron ◽  
Joanne Keenan ◽  
...  
Keyword(s):  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Molecular Basis ◽  
Chinese Hamster ◽  
Proteome Analysis ◽  
Cho Cell ◽  
Related Phenotype

Efficient Selection of Stable Chinese Hamster Ovary (CHO) Cell Lines for Expression of Recombinant Proteins by Using Human Interferon β SAR Element

2008 ◽  
Vol 21 (3) ◽  
pp. 933-937 ◽  
Author(s):  
Jeong Do Kim ◽  
Yeup Yoon ◽  
Hye-Yeon Hwang ◽  
Jeong Soo Park ◽  
Sungryul Yu ◽  
...  
Keyword(s):  
Cell Lines ◽  
Recombinant Proteins ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Human Interferon ◽  
Interferon Β ◽  
Cho Cell ◽  
Efficient Selection ◽  
Selection Of

scholarly journals Evidence for intracellular partitioning of serine and glycine metabolism in Chinese hamster ovary cells

1996 ◽  
Vol 313 (3) ◽  
pp. 991-996 ◽  
Author(s):  
Michael R. NARKEWICZ ◽  
S. David SAULS ◽  
Susan S. TJOA ◽  
Cecilia TENG ◽  
Paul V. FENNESSEY
Keyword(s):  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Chinese Hamster Ovary Cells ◽  
Mitochondrial Protein ◽  
Chinese Hamster ◽  
Serine Hydroxymethyltransferase ◽  
Cho Cell ◽  
Ovary Cells ◽  
Glycine Metabolism

Serine hydroxymethyltransferase (SHMT) is the primary enzyme in the interconversion of serine and glycine. The roles of mitochondrial and cytosolic SHMT in the interconversion of serine and glycine were determined in two Chinese hamster ovary (CHO) cell lines that both contain cytosolic SHMT but either have (CHOm+) or lack (CHOm-) mitochondrial SHMT. Mitochondrial SHMT activity was significantly reduced in CHOm- (0.24±0.11 nmol/min per mg of mitochondrial protein) compared with CHOm+ (3.21±0.66 nmol/min per mg of mitochondrial protein; P = 0.02) cells, whereas cytosolic SHMT activity was similar in CHOm- and CHOm+ cells (1.09±0.31 and 1.53±0.12 nmol/min per mg of cytosolic protein respectively; P = 0.57). In CHOm+ and CHOm- cells, the relative flux of glycine to serine measured with either [1-13C]- or [2-13C]-glycine was similar (CHOm-: 538±82 nmol/24 per mg of DNA; CHOm+: 616±88 nmol/24 h per mg of DNA; P = 0.42). In contrast, the relative flux of serine to glycine measured with [1-13C]serine was low in CHOm- cells (80±28 nmol/24 h per mg of DNA) compared with CHOm+ cells (3080±320 nmol/24 h per mg of DNA; P = 0.0001). The rate of glycine production determined by UA-2[1-13C]glycine dilution was lower in CHOm- (1200±200 nmol/24 h per mg of DNA) than CHOm+ (10200±1800 nmol/24 h per mg of DNA; P = 0.03) cells, whereas glycine utilization was similar in the two cell lines. Serine production was similar in the two cell lines but serine utilization was lower in CHOm- (3800±1200 μmol/24 h per mg of DNA) than CHOm+ (6600±1000 nmol/24 h per mg of DNA; P = 0.0002) cells. Increasing the serine concentration in the medium resulted in an increase in glycine production in CHOm+ but not in CHOm- cells. Intracellular studies with [1-13C]serine confirm the findings of decreased glycine production from serine. In CHO cells there is partitioning of intracellular serine and glycine metabolism. Our data support the hypothesis that mitochondrial SHMT is the primary pathway for serine into glycine interconversion.

scholarly journals Restoration of deficient DNA Repair Genes Mitigates Genome Instability and Increases Productivity of Chinese Hamster Ovary Cells

2021 ◽  
Author(s):  
Philipp N. Spahn ◽  
Xiaolin Zhang ◽  
Qing Hu ◽  
Nathaniel K. Hamaker ◽  
Hooman Hefzi ◽  
...  
Keyword(s):  
Dna Repair ◽  
Cell Line ◽  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Genome Instability ◽  
Therapeutic Proteins ◽  
Chinese Hamster ◽  
Dna Repair Genes ◽  
Cho Cell ◽  
Repair Genes

AbstractChinese Hamster Ovary (CHO) cells are the primary host used for manufacturing of therapeutic proteins. However, production instability of high-titer cell lines is a major problem and is associated with genome instability, as chromosomal aberrations reduce transgene copy number and decrease protein titer. We analyzed whole-genome sequencing data from 11 CHO cell lines and found deleterious single-nucleotide polymorphisms (SNPs) in DNA repair genes. Comparison with other mammalian cells confirmed DNA repair is compromised in CHO. Restoration of key DNA repair genes by SNP reversal or expression of intact cDNAs improved DNA repair and genome stability. Moreover, the restoration of LIG4 and XRCC6 in a CHO cell line expressing secreted alkaline phosphatase mitigated transgene copy loss and improved protein titer retention. These results show for the first time that correction of key DNA repair genes yields considerable improvements in stability and protein expression in CHO, and provide new opportunities for cell line development and a more efficient and sustainable production of therapeutic proteins.

scholarly journals Evidence for co-translational misincorporation of non-canonical amino acid hydroxyproline in recombinant antibodies produced in Chinese Hamster Ovary (CHO) cell lines

PLoS ONE ◽  
2020 ◽  
Vol 15 (10) ◽  
pp. e0241250
Author(s):  
Shanta Boddapati ◽  
Jason Gilmore ◽  
Kyle Boone ◽  
John Bushey ◽  
Jonathan Ross ◽  
...  
Keyword(s):  
Amino Acid ◽  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Recombinant Antibodies ◽  
Cho Cell ◽  
Canonical Amino Acid

scholarly journals Mechanisms underlying epigenetic and transcriptional heterogeneity in Chinese hamster ovary (CHO) cell lines

2016 ◽  
Vol 16 (1) ◽  
Author(s):  
Nathalie Veith ◽  
Holger Ziehr ◽  
Roderick A. F. MacLeod ◽  
Stella Marie Reamon-Buettner
Keyword(s):  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Cho Cell

ERCC1 mutations in UV-sensitive Chinese hamster ovary (CHO) cell lines

1998 ◽  
Vol 407 (3) ◽  
pp. 269-276 ◽  
Author(s):  
Tsuyuko Hayashi ◽  
Masashi Takao ◽  
Kiyoji Tanaka ◽  
Akira Yasui
Keyword(s):  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Chinese Hamster ◽  
Cho Cell

scholarly journals Using Metabolomics to Identify Cell Line-Independent Indicators of Growth Inhibition for Chinese Hamster Ovary Cell-Based Bioprocesses

Metabolites ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 199 ◽  
Author(s):  
Nicholas Alden ◽  
Ravali Raju ◽  
Kyle McElearney ◽  
James Lambropoulos ◽  
Rashmi Kshirsagar ◽  
...  
Keyword(s):  
Growth Inhibition ◽  
Cell Line ◽  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Cho Cells ◽  
Feeding Strategy ◽  
Chinese Hamster ◽  
Ovary Cell ◽  
Growth Stages ◽  
Cho Cell

Chinese hamster ovary (CHO) cells are widely used for the production of biopharmaceuticals. Efforts to improve productivity through medium design and feeding strategy optimization have focused on preventing the depletion of essential nutrients and managing the accumulation of lactate and ammonia. In addition to ammonia and lactate, many other metabolites accumulate in CHO cell cultures, although their effects remain largely unknown. Elucidating these effects has the potential to further improve the productivity of CHO cell-based bioprocesses. This study used untargeted metabolomics to identify metabolites that accumulate in fed-batch cultures of monoclonal antibody (mAb) producing CHO cells. The metabolomics experiments profiled six cell lines that are derived from two different hosts, produce different mAbs, and exhibit different growth profiles. Comparing the cell lines’ metabolite profiles at different growth stages, we found a strong negative correlation between peak viable cell density (VCD) and a tryptophan metabolite, putatively identified as 5-hydroxyindoleacetaldehyde (5-HIAAld). Amino acid supplementation experiments showed strong growth inhibition of all cell lines by excess tryptophan, which correlated with the accumulation of 5-HIAAld in the culture medium. Prospectively, the approach presented in this study could be used to identify cell line- and host-independent metabolite markers for clone selection and bioprocess development.

scholarly journals Restoration of deficient DNA Repair Genes Mitigates Genome Instability and Increases Productivity of Chinese Hamster Ovary Cells

2021 ◽  
Author(s):  
Philipp Spahn ◽  
Xiaolin Zhang ◽  
Qing Hu ◽  
Nathaniel Hamaker ◽  
Hooman Hefzi ◽  
...  
Keyword(s):  
Dna Repair ◽  
Cell Line ◽  
Cell Lines ◽  
Chinese Hamster Ovary ◽  
Genome Instability ◽  
Therapeutic Proteins ◽  
Chinese Hamster ◽  
Dna Repair Genes ◽  
Cho Cell ◽  
Repair Genes

Abstract Chinese Hamster Ovary (CHO) cells are the primary host used for manufacturing of therapeutic proteins. However, production instability of high-titer cell lines is a major problem and is associated with genome instability, as chromosomal aberrations reduce transgene copy number and decrease protein titer. We analyzed whole-genome sequencing data from 11 CHO cell lines and found deleterious single-nucleotide polymorphisms (SNPs) in DNA repair genes. Comparison with other mammalian cells confirmed DNA repair is compromised in CHO. Restoration of key DNA repair genes by SNP reversal or expression of intact cDNAs improved DNA repair and genome stability. Moreover, the restoration of LIG4 and XRCC6 in a CHO cell line expressing secreted alkaline phosphatase mitigated transgene copy loss and improved protein titer retention. These results show for the first time that correction of key DNA repair genes yields considerable improvements in stability and protein expression in CHO, and provide new opportunities for cell line development and a more efficient and sustainable production of therapeutic proteins.

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