Compositions and methods for humanization and optimization...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/54 (2006.01) A01H 5/00 (2006.01) C12N 5/10 (2006.01) C12N 9/10 (2006.01) C12N 15/11 (2006.01) C12N 15/82 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2637252

Methods for altering the N-glycosylation pattern of proteins in higher plants are provided. The methods comprise introducing into the plant a recombinant construct that provides for the inhibition of expression of .alpha.l,3- fucosyltransferase (FucT) and .beta. 1 ,2-xylosyltransferase (XyIT) in a plant. Use of these constructs to inhibit or suppress expression of both of these enzymes, and isoforms thereof, advantageously provides for the production of endogenous and heterologous proteins having a "humanized" N- glycosylation pattern without impacting plant growth and development. Stably transformed higher plants having this protein N-glycosylation pattern are provided. Glycoprotein compositions, including monoclonal antibody compositions, having substantially homogeneous glycosylation profiles, and which are substantially homogeneous for the GO glycoform, are also provided.

L'invention concerne des procédés qui permettent de modifier le motif de N-glycosylation des protéines chez les plantes supérieures. Les procédés de l'invention consistent à introduire dans la plante un produit de recombinaison qui inhibe l'expression de l'.alpha.l,3-fucosyltransférase (FucT) et de la .beta. 1 ,2-xylosyltransférase (XyIT) chez une plante. L'utilisation de produits de recombinaison de ce type pour inhiber ou supprimer l'expression des deux enzymes précitées, et de leur isoformes, offre l'avantage de permettre la production de protéines endogènes et hétérologues présentant un motif de N-glycosylation "humanisé" sans entraver la croissance et le développement des plantes. L'invention porte sur des plantes supérieures transformées de manière stable qui renferment ledit motif de N-glycosylation des protéines. L'invention se rapporte également à des compositions de glycoprotéines, y compris des compositions d'anticorps monoclonaux, possédant des profils de glycosylation sensiblement homogènes pour la glycoforme GO.

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