Transient Glyco-Engineering to Produce Recombinant IgA1 with Defined N- and O-Glycans in Plants.

Autor:	Dicker M; Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Austria., Tschofen M; Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Austria., Maresch D; Department of Chemistry, University of Natural Resources and Life Sciences Vienna, Austria., König J; Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Austria., Juarez P; Institute of Molecular and Cellular Plant Biology, Spanish Research Council Agency - Polytechnic University of Valencia Valencia, Spain., Orzaez D; Institute of Molecular and Cellular Plant Biology, Spanish Research Council Agency - Polytechnic University of Valencia Valencia, Spain., Altmann F; Department of Chemistry, University of Natural Resources and Life Sciences Vienna, Austria., Steinkellner H; Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Austria., Strasser R; Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Austria.
Jazyk:	angličtina
Zdroj:	Frontiers in plant science [Front Plant Sci] 2016 Jan 29; Vol. 7, pp. 18. Date of Electronic Publication: 2016 Jan 29 (Print Publication: 2016).
DOI:	10.3389/fpls.2016.00018
Abstrakt:	The production of therapeutic antibodies to combat pathogens and treat diseases, such as cancer is of great interest for the biotechnology industry. The recent development of plant-based expression systems has demonstrated that plants are well-suited for the production of recombinant monoclonal antibodies with defined glycosylation. Compared to immunoglobulin G (IgG), less effort has been undertaken to express immunoglobulin A (IgA), which is the most prevalent antibody class at mucosal sites and a promising candidate for novel recombinant biopharmaceuticals with enhanced anti-tumor activity. Here, we transiently expressed recombinant human IgA1 against the VP8* rotavirus antigen in glyco-engineered ΔXT/FT Nicotiana benthamiana plants. Mass spectrometric analysis of IgA1 glycopeptides revealed the presence of complex biantennary N-glycans with terminal N-acetylglucosamine present on the N-glycosylation site of the CH2 domain in the IgA1 alpha chain. Analysis of the peptide carrying nine potential O-glycosylation sites in the IgA1 alpha chain hinge region showed the presence of plant-specific modifications including hydroxyproline formation and the attachment of pentoses. By co-expression of enzymes required for initiation and elongation of human O-glycosylation it was possible to generate disialylated mucin-type core 1 O-glycans on plant-produced IgA1. Our data demonstrate that ΔXT/FT N. benthamiana plants can be engineered toward the production of recombinant IgA1 with defined human-type N- and O-linked glycans.
Databáze:	MEDLINE
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