Abstract
Yeast is capable of performing posttranslational modifications, such as N- or O-glycosylation. It has been demonstrated that N-glycans play critical biological roles in therapeutic glycoproteins by modulating pharmacokinetics and pharmacodynamics. However, N-glycan sites on recombinant glycoproteins produced in yeast can be underglycosylated, and hence, not completely occupied. Genomic homology analysis indicates that the Pichia pastoris oligosaccharyltransferase (OST) complex consists of multiple subunits, including OST1, OST2, OST3, OST4, OST5, OST6, STT3, SWP1, and WBP1. Monoclonal antibodies produced in P. pastoris show that N-glycan site occupancy ranges from 75-85 % and is affected mainly by the OST function, and in part, by process conditions. In this study, we demonstrate that N-glycan site occupancy of antibodies can be improved to greater than 99 %, comparable to that of antibodies produced in mammalian cells (CHO), by overexpressing Leishmania major STT3D (LmSTT3D) under the control of an inducible alcohol oxidase 1 (AOX1) promoter. N-glycan site occupancy of non-antibody glycoproteins such as recombinant human granulocyte macrophage colony-stimulating factor (rhGM-CSF) was also significantly improved, suggesting tha...Continue Reading
References
Nov 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·A H HorwitzR R Robinson
Jun 24, 1987·Biochimica Et Biophysica Acta·H A KaplanW J Lennarz
Jun 16, 1995·The Journal of Biological Chemistry·L KasturiS H Shakin-Eshleman
Feb 20, 1997·Gene·H R WaterhamJ M Cregg
Jan 24, 1998·The Journal of Biological Chemistry·D KaraogluR Gilmore
Jun 6, 1998·Biochemistry·J L MellquistS H Shakin-Eshleman
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Jan 12, 2001·Journal of Molecular Biology·A KroghE L Sonnhammer
Oct 3, 2002·The Journal of Biological Chemistry·Qi Yan, William J Lennarz
Apr 19, 2003·Proceedings of the National Academy of Sciences of the United States of America·Byung-Kwon ChoiTillman U Gerngross
May 21, 2003·The Journal of Cell Biology·IngMarie NilssonArthur E Johnson
Jun 26, 2003·Nucleic Acids Research·Ramu ChennaJulie D Thompson
Jun 26, 2003·Protein Science : a Publication of the Protein Society·Jing Liu, Arcady Mushegian
Aug 30, 2003·Science·Stephen R HamiltonTillman U Gerngross
Sep 30, 2003·Glycobiology·Shifra Ben-DorNathan Sharon
May 7, 2004·Applied and Environmental Microbiology·Michael WardHuaming Wang
Jun 17, 2005·Journal of Pharmaceutical Sciences·Angus M Sinclair, Steve Elliott
Aug 13, 2005·Glycobiology·Aixin Yan, William J Lennarz
Aug 30, 2005·Biochimica Et Biophysica Acta·Jullian JonesMichael J Betenbaugh
Jan 24, 2006·Nature Biotechnology·Huijuan LiTillman U Gerngross
Sep 9, 2006·Science·Stephen R HamiltonTillman U Gerngross
Oct 24, 2007·Current Opinion in Biotechnology·Stephen R Hamilton, Tillman U Gerngross
Nov 30, 2007·The EMBO Journal·Mayumi IguraDaisuke Kohda
Mar 28, 2008·Glycoconjugate Journal·Byung-Kwon ChoiStefan Wildt
Jul 4, 2008·Molecular Biology of the Cell·Farnoush Parsaie NasabMarkus Aebi
Oct 29, 2008·Glycobiology·Katrin HeseLudwig Lehle
Jan 10, 2009·Nature Protocols·Pieter P JacobsNico Callewaert
Jan 24, 2009·Journal of Biotechnology·Thomas I PotgieterMarc d'Anjou
May 7, 2009·Biotechnology and Bioengineering·Martin GawlitzekRobert Kiss
Jun 25, 2009·Proceedings of the National Academy of Sciences of the United States of America·Benjamin L SchulzMarkus Aebi
Sep 22, 2009·Methods in Molecular Biology·Theresa C SwayneLiza A Pon
Dec 17, 2009·The Journal of Biological Chemistry·Nobuo MaitaDaisuke Kohda
Aug 17, 2010·Journal of Industrial Microbiology & Biotechnology·Gavin C BarnardPiotr Bobrowicz
Nov 16, 2010·Protein Expression and Purification·Youwei JiangHuijuan Li
Mar 23, 2011·Glycobiology·Andrew M GoetzeGregory C Flynn
Apr 14, 2011·MAbs·Ningyan ZhangDongxing Zha
Nov 22, 2011·Journal of Biotechnology·Juergen H NettNatarajan Sethuraman
Citations
Sep 3, 2013·Applied Biochemistry and Biotechnology·Renata DamianiMaria Teresa C P Ribela
Dec 4, 2012·Applied and Environmental Microbiology·Farnoush Parsaie NasabAlexander Daniel Frey
May 15, 2013·PloS One·Rebecca ArgyrosBo Jiang
Feb 15, 2014·Applied Microbiology and Biotechnology·Sujatha Gomathinayagam, Stephen R Hamilton
Feb 15, 2014·Applied Microbiology and Biotechnology·Daniel HopkinsStephen R Hamilton
Jul 16, 2015·Expert Opinion on Biological Therapy·Martina Dicker, Richard Strasser
Feb 13, 2016·Journal of Pharmaceutical Sciences·Solomon Z OkbazghiThomas J Tolbert
Sep 5, 2015·Current Opinion in Biotechnology·Diana Chinyere Anyaogu, Uffe Hasbro Mortensen
Dec 15, 2015·Microbial Cell Factories·Rachel Chen
Mar 15, 2016·Applied Biochemistry and Biotechnology·Weiwei ChenLidan Ye
Nov 11, 2014·Applied Microbiology and Biotechnology·Daniel HopkinsStephen R Hamilton
Feb 28, 2015·Applied Microbiology and Biotechnology·Verena PuxbaumBrigitte Gasser
Dec 10, 2014·Glycobiology·Alexander D Frey, Markus Aebi
Jan 20, 2015·Future Microbiology·Bram LaukensNico Callewaert
Nov 12, 2016·World Journal of Microbiology & Biotechnology·Amjad Hayat KhanAzam Rahimpour
Jan 24, 2015·Scientific Reports·Krystal Teasley HamorskyNobuyuki Matoba
Jul 12, 2014·Current Opinion in Biotechnology·Michael A Meehl, Terrance A Stadheim
Feb 27, 2018·Plant Biotechnology Journal·Alexandra CastilhoRichard Strasser
Sep 21, 2017·The Journal of Biological Chemistry·Anders JinnelovMichael A J Ferguson
Sep 10, 2014·Biotechnology Progress·Muralidhar MallemByung-Kwon Choi
Nov 21, 2018·Biotechnology Progress·Amjad Hayat Khan, Rahmah Noordin
Oct 23, 2016·Microbial Cell Factories·Sha XuXiao-Dong Gao
Aug 5, 2020·Molecular Biotechnology·Amjad Hayat KhanRahmah Noordin
Dec 8, 2015·Glycoconjugate Journal·Lucia F Zacchi, Benjamin L Schulz
Jun 27, 2019·Biotechnology and Bioengineering·Thomas AmannMikael Rørdam Andersen
Feb 6, 2020·FEMS Yeast Research·Eun Jung ThakHyun Ah Kang
Oct 17, 2017·Molecular Immunology·Ishan S ShahThomas J Tolbert
Jun 12, 2020·ACS Synthetic Biology·Weston KightlingerMichael C Jewett
Mar 19, 2014·New Biotechnology·Mari A PiirainenAlexander D Frey