Improving the production of recombinant human bone morphogenetic protein-4 in Chinese hamster ovary cell cultures by inhibition of undesirable endocytosis

Biotechnology and Bioengineering
Che Lin KimGyun Min Lee

Abstract

Endocytic regulation serves a critical role in modulating the extracellular level of signaling molecules, such as bone morphogenetic proteins (BMPs). Unfortunately, endocytosis may result in poor yields of recombinant human BMP-4 (rhBMP-4) from Chinese hamster ovary (CHO) cell cultures. When rhBMP-4 was incubated with CHO cells, rhBMP-4 was actively internalized into cells. Cell surface bound heparan sulfate proteoglycans (HSPGs) served as the major receptors for rhBMP-4 internalization. Removal of cell surface heparan sulfate (HS) by heparinases or reduction of HSPG synthesis by knockdown of xylosyltransferase2 (xylt2) in CHO cells decreased internalization of rhBMP-4. In addition, treatment with endocytosis inhibitors (chlorpromazine, genistein, and dynasore) identified a clathrin- and dynamin-dependent endocytic pathway as the major route for rhBMP-4 internalization. To enhance product yield by minimizing rhBMP-4 internalization in recombinant CHO (rCHO) cell cultures, we have tested various strategies to reduce HSPG synthesis (knockdown of xylt2 and sodium chlorate treatment) or inhibit the binding of rhBMP-4 to cell-surface-bound HSPGs (supplementation with heparin or dextran sulfate [DS]). Among these approaches, DS, whic...Continue Reading

References

Dec 15, 1986·Biochemical and Biophysical Research Communications·P A Baeuerle, W B Huttner
Jun 7, 2002·Genes & Development·Elaine S SetoThomas E Lloyd
Aug 13, 2003·The Journal of Biological Chemistry·Takatora TakadaRyutaro Kamijo
Aug 30, 2005·Biochimica Et Biophysica Acta·Lucas Pelkmans
Aug 23, 2006·Molecular and Cellular Biology·Anke HartungPetra Knaus
Oct 6, 2006·The Journal of Biological Chemistry·Xiangyang JiaoDavid L Glaser
Nov 10, 2006·Proceedings of the National Academy of Sciences of the United States of America·A Jamila NewtonVenkatesh N Murthy
Aug 8, 2007·Biotechnology and Bioengineering·Sun Ok Hwang, Gyun Min Lee
Apr 4, 2008·American Journal of Physiology. Cell Physiology·Shaukat A KhanPankaj Gupta
Mar 26, 2009·Annual Review of Biochemistry·Gary J Doherty, Harvey T McMahon
Jul 25, 2009·Applied Microbiology and Biotechnology·Hiroyuki MukaiyamaKaoru Takegawa
Nov 10, 2009·Cytokine & Growth Factor Reviews·Christina SieberPetra Knaus
Oct 21, 2010·Cellular Signalling·Beth BragdonAnja Nohe
Dec 15, 2010·European Cells & Materials·Yaiza Lópiz-MoralesFernando Marco
Mar 12, 2011·Growth Factors·Sabrina von EinemElisabeth Schwarz
Oct 19, 2012·Journal of Cell Science·Hamed AlborziniaStefan Wölfl
Nov 8, 2012·Development·Thomas E WillnowAnnette Hammes
Feb 5, 2013·Journal of Biotechnology·Madhavi SathyamurthyGyun Min Lee
Jun 1, 2013·Clinical Journal of the American Society of Nephrology : CJASN·Jonathan L MoskowitzPeter Kotanko
Jul 31, 2013·Proceedings of the National Academy of Sciences of the United States of America·Brandon B HolmesMarc I Diamond
Nov 5, 2013·Cold Spring Harbor Perspectives in Biology·Linton M Traub, Juan S Bonifacino
Jan 7, 2014·Journal of Dental Research·A C CarreiraJ M Granjeiro
Nov 8, 2014·The Biochemical Journal·Rachel H ChurchDerek P Brazil

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