Main content

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

Show full item record

Title: Generation of Nicotiana benthamiana with humanized N-glycosylation for therapeutic monoclonal antibody production
Author: So, Kenny Kin Yang
Department: School of Environmental Sciences
Program: Environmental Sciences
Advisor: Hall, J. Christopher
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.
URI: http://hdl.handle.net/10214/8918
Date: 2015-06
Rights: Attribution-NonCommercial-NoDerivs 2.5 Canada
Terms of Use: All items in the Atrium are protected by copyright with all rights reserved unless otherwise indicated.


Files in this item

Files Size Format View Description
So_Kenny_201506_MSc.pdf 2.338Mb PDF View/Open So_Kenny_201506_MSc

This item appears in the following Collection(s)

Show full item record

Attribution-NonCommercial-NoDerivs 2.5 Canada Except where otherwise noted, this item's license is described as Attribution-NonCommercial-NoDerivs 2.5 Canada