Transient glyco-engineering to produce recombinant IgA1 with defined N-and O-glycans in plants
| Autor: | Dicker, Martina, Tschofen, Marc, Maresch, Daniel, König, Julia, Juarez, Paloma, Orzaez, Diego, Altmann, Friedrich, Steinkellner, Herta, Strasser, Richard |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
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| Zdroj: | RiuNet. Repositorio Institucional de la Universitat Politécnica de Valéncia instname Frontiers in Plant Science |
| Popis: | [EN] 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 XT/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 XT/FT N. benthamiana plants can be engineered toward the production of recombinant IgA1 with defined human-type Nand O-linked glycans. This work was supported by a grant from the Austrian Federal Ministry of Transport, Innovation and Technology (bmvit) and Austrian Science Fund (FWF): TRP 242-B20 and by the Austrian Research Promotion Agency (Laura Bassi Center of Expertise "Plant produced Bio-Pharmaceuticals" Grant Nr. 822757). |
| Databáze: | OpenAIRE |
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