A novel baculovirus vector for the production of nonfucosylated recombinant glycoproteins in insect cells

Glycobiology
Hideaki Mabashi-AsazumaDonald L Jarvis

Abstract

Glycosylation is an important attribute of baculovirus-insect cell expression systems, but some insect cell lines produce core α1,3-fucosylated N-glycans, which are highly immunogenic and render recombinant glycoproteins unsuitable for human use. To address this problem, we exploited a bacterial enzyme, guanosine-5'-diphospho (GDP)-4-dehydro-6-deoxy-d-mannose reductase (Rmd), which consumes the GDP-l-fucose precursor. We expected this enzyme to block glycoprotein fucosylation by blocking the production of GDP-l-fucose, the donor substrate required for this process. Initially, we engineered two different insect cell lines to constitutively express Rmd and isolated subclones with fucosylation-negative phenotypes. However, we found the fucosylation-negative phenotypes induced by Rmd expression were unstable, indicating that this host cell engineering approach is ineffective in insect systems. Thus, we constructed a baculovirus vector designed to express Rmd immediately after infection and facilitate the insertion of genes encoding any glycoprotein of interest for expression later after infection. We used this vector to produce a daughter encoding rituximab and found, in contrast to an Rmd-negative control, that insect cells infect...Continue Reading

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Citations

Jul 16, 2015·Expert Opinion on Biological Therapy·Martina Dicker, Richard Strasser
Jul 15, 2015·Insect Biochemistry and Molecular Biology·Hideaki Mabashi-AsazumaDonald L Jarvis
Aug 5, 2015·ACS Chemical Biology·Hideaki Mabashi-AsazumaDonald L Jarvis
Sep 24, 2014·The Journal of General Virology·Monique M van OersJust M Vlak
Jun 29, 2014·Biochemical and Biophysical Research Communications·Hideyuki IharaYoshitaka Ikeda
Nov 12, 2016·World Journal of Microbiology & Biotechnology·Amjad Hayat KhanAzam Rahimpour
Jul 15, 2020·Biotechnology and Bioengineering·Ajay B MaghodiaDonald L Jarvis
Mar 28, 2019·Annual Review of Biochemistry·Lai-Xi WangTiezheng Li
Jun 12, 2020·ACS Synthetic Biology·Weston KightlingerMichael C Jewett

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