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Augmenting glycosylation-directed folding pathways enhances the fidelity of HIV Env immunogen production in plants.
Margolin, Emmanuel; Allen, Joel D; Verbeek, Matthew; Chapman, Ros; Meyers, Ann; van Diepen, Michiel; Ximba, Phindile; Motlou, Thopisang; Moore, Penny L; Woodward, Jeremy; Strasser, Richard; Crispin, Max; Williamson, Anna-Lise; Rybicki, Edward P.
Afiliação
  • Margolin E; Department of Pathology, Division of Medical Virology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Allen JD; Wellcome Trust Centre for Infectious Disease Research in Africa, University of Cape Town, Cape Town, South Africa.
  • Verbeek M; Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Chapman R; Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa.
  • Meyers A; School of Biological Sciences, University of Southampton, Southampton, UK.
  • van Diepen M; Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa.
  • Ximba P; Department of Pathology, Division of Medical Virology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Motlou T; Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Moore PL; Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa.
  • Woodward J; Department of Pathology, Division of Medical Virology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Strasser R; Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Crispin M; Department of Pathology, Division of Medical Virology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Williamson AL; Institute of Infectious Disease and Molecular Medicine, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.
  • Rybicki EP; National Institute for Communicable Diseases of the National Health Laboratory Service, Centre for HIV and STIs, Johannesburg, South Africa.
Biotechnol Bioeng ; 119(10): 2919-2937, 2022 10.
Article em En | MEDLINE | ID: mdl-35781691
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 α1,3-fucosyltransferase and ß1,2-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 the mammalian protein. Immunized rabbits developed comparable immune responses to the plant-produced and mammalian cell-derived antigens, including the induction of autologous neutralizing antibodies when the proteins were used to boost DNA and modified vaccinia Ankara virus-vectored vaccines. This study demonstrates that engineering glycosylation-directed folding offers a promising route to enhance the production of complex viral glycoproteins in plants.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Infecções por HIV / Anticorpos Neutralizantes Limite: Animals Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Infecções por HIV / Anticorpos Neutralizantes Limite: Animals Idioma: En Ano de publicação: 2022 Tipo de documento: Article