Augmenting glycosylation-directed folding pathways enhances the fidelity of HIV Env immunogen production in plants

Augmenting glycosylation-directed folding pathways enhances the fidelity of HIV Env immunogen production in plants. Margolin, E., Allen, J. D, Verbeek, M., Chapman, R., Meyers, A., Michiel Van Diepen, |, Ximba, P., Motlou, T., Penny, |, Moore, L, Woodward, J., Strasser, | R., Crispin, M., Williamson, A., & Rybicki, E. P Biotechnology and Bioengineering, John Wiley & Sons, Ltd, jul, 2022.

Paper doi abstract bibtex

Heterologous glycoprotein production relies on host glycosylation-dependent folding. When the biosynthetic machinery differs from the usual expression host, there is scope to remodel the assembly pathway to enhance glycoprotein production. Here we explore the integration of chaperone coexpression with glyco-engineering to improve the production of a model HIV-1 envelope antigen. Calreticulin was coexpressed to support protein folding together with Leishmania major STT3D oligosaccharyltransferase, to improve glycan occupancy, RNA interference to suppress the formation of truncated glycans, and Nicotiana benthamiana plants lacking $α$1,3-fucosyltransferase and $β$1,2-xylosyltransferase was used as an expression host to prevent plant-specific complex N-glycans forming. This approach reduced the formation of undesired aggregates, which predominated in the absence of glyco-engineering. The resulting antigen also exhibited increased glycan occupancy, albeit to a slightly lower level than the equivalent mammalian cell-produced protein. The antigen was decorated almost exclusively with oligomannose glycans, which were less processed compared with Biotechnol Bioeng. 2022;1-19. wileyonlinelibrary.com/journal/bit | 1 This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

@article{Margolin2022a,
abstract = {Heterologous glycoprotein production relies on host glycosylation-dependent folding. When the biosynthetic machinery differs from the usual expression host, there is scope to remodel the assembly pathway to enhance glycoprotein production. Here we explore the integration of chaperone coexpression with glyco-engineering to improve the production of a model HIV-1 envelope antigen. Calreticulin was coexpressed to support protein folding together with Leishmania major STT3D oligosaccharyltransferase, to improve glycan occupancy, RNA interference to suppress the formation of truncated glycans, and Nicotiana benthamiana plants lacking $\alpha$1,3-fucosyltransferase and $\beta$1,2-xylosyltransferase was used as an expression host to prevent plant-specific complex N-glycans forming. This approach reduced the formation of undesired aggregates, which predominated in the absence of glyco-engineering. The resulting antigen also exhibited increased glycan occupancy, albeit to a slightly lower level than the equivalent mammalian cell-produced protein. The antigen was decorated almost exclusively with oligomannose glycans, which were less processed compared with Biotechnol Bioeng. 2022;1-19. wileyonlinelibrary.com/journal/bit | 1 This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.},
author = {Margolin, Emmanuel and Allen, Joel D and Verbeek, Matthew and Chapman, Ros and Meyers, Ann and {Michiel Van Diepen}, | and Ximba, Phindile and Motlou, Thopisang and Penny, | and Moore, L and Woodward, Jeremy and Strasser, | Richard and Crispin, Max and Williamson, Anna-Lise and Rybicki, Edward P},
doi = {10.1002/BIT.28169},
file = {:C$\backslash$:/Users/01462563/AppData/Local/Mendeley Ltd./Mendeley Desktop/Downloaded/Margolin et al. - 2022 - Augmenting glycosylation-directed folding pathways enhances the fidelity of HIV Env immunogen production in pla.pdf:pdf},
issn = {1097-0290},
journal = {Biotechnology and Bioengineering},
keywords = {OA,chaperones,engineering,fund{\_}ack,glyco,glycosylation,immunogenicity,neutralization,occupancy,original,vaccine},
mendeley-tags = {OA,fund{\_}ack,original},
month = {jul},
pages = {10.1002/bit.28169},
publisher = {John Wiley {\&} Sons, Ltd},
title = {{Augmenting glycosylation-directed folding pathways enhances the fidelity of HIV Env immunogen production in plants}},
url = {https://onlinelibrary.wiley.com/doi/full/10.1002/bit.28169 https://onlinelibrary.wiley.com/doi/abs/10.1002/bit.28169 https://onlinelibrary.wiley.com/doi/10.1002/bit.28169},
year = {2022}
}

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