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Generation of Nicotiana benthamiana with humanized N-glycosylation for therapeutic monoclonal antibody production

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dc.contributor.advisor Hall, J. Christopher So, Kenny Kin Yang 2015-06-12T19:27:17Z 2015-06-12T19:27:17Z 2015-06 2015-06-10 2015-06-12
dc.description.abstract To use plants for therapeutic monoclonal antibody (mAb) production, their N-glycosylation pathway must be humanized by precluding the attachment of the human-immunogenic sugars α1,3-fucose and β1,2-xylose, followed by the attachment of β1,4-galactose. A Nicotiana benthamiana line with humanized N-glycosylation was generated by transforming an α1,3-fucosylation and β1,2-xylosylation knockdown line (ΔFX) with the coding sequence (CDS) of a chimeric human β1,4-galactosyltransferase (hGalT) enzyme. The mAb trastuzumab, produced in this line, was galactosylated indicating this line to be suitable for therapeutic mAb production. Two new α1,3-fucosylation and β1,2-xylosylation deficient lines were also created. The first line, generated using RNA interference targeting the α1,3-fucosyltransferase and β1,2-xylosyltransferase genes, showed 55% knockdown of α1,3-fucosylation and β1,2-xylosation. The second line, achieved by transforming the chimeric hGalT CDS into wildtype N. benthamiana, showed knockdown of α1,3-fucosylation and β1,2-xylosylation comparable to the ΔFX line. These two knockdown lines can be used for producing non-galactosylated protein pharmaceuticals. en_US
dc.language.iso en en_US
dc.rights Attribution-NonCommercial-NoDerivs 2.5 Canada *
dc.rights.uri *
dc.subject recombinant protein en_US
dc.subject N-glycosylation en_US
dc.subject monoclonal antibody en_US
dc.subject Agrobacterium-mediated transformation en_US
dc.subject trastuzumab en_US
dc.subject Nicotiana benthamiana en_US
dc.title Generation of Nicotiana benthamiana with humanized N-glycosylation for therapeutic monoclonal antibody production en_US
dc.type Thesis en_US Environmental Sciences en_US Master of Science en_US School of Environmental Sciences en_US
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Attribution-NonCommercial-NoDerivs 2.5 Canada Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 2.5 Canada