Comparative characterization of the glycosylation profiles of an influenza hemagglutinin produced in plant and insect hosts.

Proteomics
Sheng ZhangTheodore W Thannhauser

Abstract

The main objective of this study was to characterize the N-linked glycosylation profiles of recombinant hemagglutinin (HA) proteins expressed in either insect or plant hosts, and to develop a mass spectrometry based workflow that can be used in quality control to assess batch-to-batch reproducibility for recombinant HA glycosylation. HA is a surface glycoprotein of the influenza virus that plays a key role in viral infectivity and pathogenesis. Characterization of the glycans for plant recombinant HA from the viral strain A/California/04/09 (H1N1) has not yet been reported. In this study, N-linked glycosylation patterns of the recombinant HAs from both insect and plant hosts were characterized by precursor ion scan-driven data-dependent analysis followed by high-resolution MS/MS analysis of the deglycosylated tryptic peptides. Five glycosylation sites (N11, N23, N276, N287, and N481) were identified containing high mannose type glycans in plant-expressed HAs, and complex type glycoforms for the insect-expressed HA. More than 95% site occupancy was observed for all glycosylation sites except N11, which was 60% occupied. Multiple-reaction monitoring based quantitation analysis was developed for each glycopeptide isoform and the q...Continue Reading

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Citations

Aug 19, 2014·Nature Chemical Biology·Anne A OllisMatthew P DeLisa
Dec 4, 2014·Virology Journal·Edward P Rybicki
Apr 14, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·John M HickeyDavid B Volkin
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Nov 28, 2017·Biotechnology and Bioengineering·Jennifer A SchoborgMichael C Jewett
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Feb 1, 2014·Molecular & Cellular Proteomics : MCP·Eliel Ruiz-MayJocelyn K C Rose
Nov 2, 2020·Biophysical Journal·Christian SeitzJ Andrew McCammon
Jan 9, 2020·Journal of Proteome Research·Celina L KeatingDaniel Lingwood

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