<oai_dc:dc xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:oai_dc="http://www.openarchives.org/OAI/2.0/oai_dc/" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd">
  <dc:title xml:lang="eng">Glycoproteomic and Single-Protein Glycomic Analyses Reveal Zwitterionic N-Glycans on Natural and Recombinant Proteins Derived From Insect Cells</dc:title>
  <dc:format>application/pdf</dc:format>
  <dc:language>eng</dc:language>
  <dc:type xml:lang="eng">Text</dc:type>
  <dc:type xml:lang="eng">journal article</dc:type>
  <dc:type xml:lang="ita">Testo</dc:type>
  <dc:type xml:lang="ita">Articolo di rivista</dc:type>
  <dc:creator>Shi Yan</dc:creator>
  <dc:creator>Jorick Vanbeselaere</dc:creator>
  <dc:creator>Callum Ives</dc:creator>
  <dc:creator>David Stenitzer</dc:creator>
  <dc:creator>Lena Nuschy</dc:creator>
  <dc:creator>Florian Woels</dc:creator>
  <dc:creator>Katharina Paschinger</dc:creator>
  <dc:creator>Elisa Fadda</dc:creator>
  <dc:creator>Johannes Stadlmann</dc:creator>
  <dc:creator>Iain Wilson</dc:creator>
  <dc:subject xml:lang="eng">Baculovirus</dc:subject>
  <dc:subject xml:lang="eng">Fucose</dc:subject>
  <dc:subject xml:lang="eng">Glucuronic Acid</dc:subject>
  <dc:subject xml:lang="eng">Insect Cells</dc:subject>
  <dc:subject xml:lang="eng">N-glycans</dc:subject>
  <dc:subject xml:lang="eng">Phosphorylcholine</dc:subject>
  <dc:description xml:lang="eng">Insect cells are a convenient cell factory to produce recombinant glycoproteins. Their glycosylation potential is believed to be simple, needing primarily addition of glycosyltransferases to humanize the recombinant products. In this study, the native glycoproteome of Spodoptera frugiperda Sf9 and Trichoplusia ni High Five cells, examined using an LC-MS/MS approach, revealed not only which proteins are N-glycosylated but also indicated that the N-glycomes contain novel glucuronylated and phosphorylcholine-modified glycans, in addition to typical oligomannosidic and fucosylated structures. These data were corroborated by a parallel MALDI-TOF MS/MS analysis of N-glycosidase-released oligosaccharides. Molecular modeling analysis of one endogenous Sf9 glycoprotein correlated the occurrence of complex and oligomannosidic N-glycans with the accessibility of the occupied N-glycosylation sites. Further, we showed that the N-glycans of influenza hemagglutinins and SARS-CoV-2 spike glycoprotein produced in Spodoptera cells possess a number of glycan structures modified with phosphorylcholine, but core difucosylation was minimal; in contrast, the Trichoplusia-produced hemagglutinin had only traces of the former type, while the latter was dominant. Detection of phosphorylcholine on these glycoproteins correlated with binding to human C-reactive protein. In conclusion, not just oligomannosidic or truncated paucimannosidic N-glycans, but structures with immunogenic features occur on both natural and recombinant glycoproteins derived from insect cell lines.</dc:description>
  <dc:type xml:lang="deu">Text</dc:type>
  <dc:type xml:lang="deu">Wissenschaftlicher Artikel</dc:type>
  <dc:publisher>American Society for Biochemistry and Molecular Biology</dc:publisher>
  <dc:rights>http://creativecommons.org/licenses/by/4.0/</dc:rights>
  <dc:rights xml:lang="eng">© 2025 The Author(s)</dc:rights>
  <dc:rights xml:lang="eng">open access</dc:rights>
  <dc:identifier>doi:10.1016/j.mcpro.2025.100981</dc:identifier>
  <dc:date>2025</dc:date>
  <dc:source xml:lang="eng">Molecular &amp; Cellular Proteomics</dc:source>
  <dc:identifier>https://phaidra.vetmeduni.ac.at/o:4833</dc:identifier>
</oai_dc:dc>