The effect of dissolved oxygen on the production and the glycosylation profile of recombinant human erythropoietin produced from CHO cells

Biotechnology and Bioengineering
Veronica RestelliM Butler

Abstract

Human recombinant erythropoietin (rHuEPO) was produced from Chinese hamster ovary (CHO) cells transfected with the human EPO gene. The cells were grown in batch cultures in controlled bioreactors in which the set-points for dissolved oxygen varied between 3% and 200%. The cell-specific growth rate and final cell yield was significantly lower under hyperoxic conditions (200% DO). However, there was no significant difference in growth rates at other oxygen levels compared to control cultures run under a normoxic condition (50% DO). The specific productivity of EPO was significantly lower at a DO set-point of 3% and 200% but maintained a consistently high value between 10% to 100% DO. The EPO produced under all conditions as analyzed by two-dimensional electrophoresis showed a molecular weight range of 33 to 37 kDa and a low isoelectric point range of 3.5 to 5.0. This corresponds to a highly glycosylated and sialylated protein with a profile showing at least seven distinct isoforms. The glycan pattern of isolated samples of EPO was analyzed by weak anion exchange (WAX) HPLC and by normal-phase HPLC incorporating sequential digestion with exoglycosidase arrays. Assigned structures were confirmed by mass spectrometry (MALDI-MS). The...Continue Reading

References

Jan 1, 1979·Methods in Enzymology·M K Patterson
Oct 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·M TakeuchiA Kobata
Jun 3, 1988·Science·J A Imlay, S Linn
Apr 11, 1987·British Dental Journal·W T MacCulloch
Jul 1, 1995·Free Radical Biology & Medicine·T D OberleyL W Oberley
May 1, 1994·Enzyme and Microbial Technology·N Jenkins, E M Curling
Oct 1, 1994·Current Opinion in Biotechnology·D C Andersen, C F Goochee
Nov 8, 1994·Proceedings of the National Academy of Sciences of the United States of America·M K ShigenagaB N Ames
Nov 1, 1996·The Journal of Biological Chemistry·M TonettiA De Flora
Aug 1, 1997·Current Opinion in Biotechnology·P M Rudd, R A Dwek
Feb 10, 1998·International Journal of Cancer. Journal International Du Cancer·K NodaN Taniguchi
May 12, 1998·Cellular and Molecular Life Sciences : CMLS·P Sears, C H Wong
Jun 17, 1998·Biotechnology Progress·J O KonzC L Cooney
Aug 1, 1996·Nature Biotechnology·N JenkinsD C James
May 6, 1999·Cellular and Molecular Life Sciences : CMLS·G Reuter, H J Gabius
Apr 4, 2000·Biotechnology and Bioengineering·M Yang, M Butler
Nov 28, 2000·International Journal of Cancer. Journal International Du Cancer·T TakahashiN Taniguchi
Jun 13, 2001·Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association·J C Egrie, J K Browne

❮ Previous
Next ❯

Citations

Jul 24, 2012·Applied Microbiology and Biotechnology·Jae Seong LeeGyun Min Lee
Mar 3, 2007·International Urology and Nephrology·A Covic, M K Kuhlmann
Jun 6, 2009·Glycobiology·Patrick HosslerZheng Jian Li
Sep 20, 2008·Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association·Adrian CovicGerard London
Jun 13, 2008·Kidney International·Sai Ram Keithi-ReddyAjay K Singh
Jun 4, 2010·Biotechnology and Bioengineering·Danny Chee Furng WongMiranda Gek Sim Yap
Nov 26, 2009·Biotechnology Progress·M Ben-TchavtchavadzeMario Jolicoeur
Jun 17, 2016·PloS One·Amanda M LewisZheng Jian Li
Oct 24, 2018·Biotechnology Journal·Viktor SandnerJarka Glassey
Dec 10, 2019·Biotechnology and Bioengineering·Valentine ChevallierLaetitia Malphettes
May 23, 2020·Scientific Reports·Kyoung SongYoung Kee Shin
Jan 8, 2021·International Journal of Molecular Sciences·Monika K NisiewiczAnna M Nowicka
Apr 21, 2021·SLAS Technology·Jonas AusterjostRickard Sjögren

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.