Glycosylated protein expression in prokaryotes

C - Chemistry – Metallurgy – 12 – N

Patent

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Details

C12N 1/21 (2006.01) C07K 14/00 (2006.01) C07K 16/00 (2006.01) C12N 15/00 (2006.01) C12P 21/00 (2006.01) C12Q 1/00 (2006.01) C12N 9/10 (2006.01) C12P 19/18 (2006.01)

Patent

CA 2711307

The invention relates to a prokaryotic host cell comprising eukaryotic glycosyttransferase activity, wherein the eukaryotic glycosyltransferase activity is eukaryotic dolichyl-linked UDP- GIcNAc transferase activity and eukaryotic mannosyltransferase activity. Also disclosed is a method of producing a glycosylated protein by providing a prokaryotic host cell comprising the eukaryotic glycosyltransferase activity and culturing the prokaryotic host cell under conditions effective to produce a glycosylated protein. Another aspect of the disclosure pertains to a method for screening bacteria or bacteriophages by expressing one or more glycans on the surface of a bacteria, attaching a label on the one or more glycans on the surface of the bacteria or a bacteriophage derived from the bacteria, and analyzing the label in a high- throughput format. A glycosylated antibody comprising an Fv portion which recognizes and binds to a native antigen and an Fc portion which is glycosylated at a conserved asparagine residue is also disclosed.

La présente invention concerne une cellule hôte procaryotique comprenant une activité glycosyltranférase eucaryotique, l'activité glycosyltranférase eucaryotique étant une activité UDP-GIcNAc dolichyl-lié transférase eucaryotique et une activité mannosyltransférase eucaryotique. Elle concerne également un procédé de production d'une protéine glycosylée comprenant le fait de fournir une cellule hôte procaryotique comprenant l'activité glycosyltranférase eucaryotique et de cultiver la cellule hôte procaryotique dans des conditions efficaces pour produire une protéine glycosylée. Un autre aspect de la présente invention concerne un procédé de criblage de bactéries ou de bactériophages par l'expression d'un ou plusieurs glycanes sur la surface d'une bactérie, l'attachement d'un marqueur sur le ou les glycanes sur la surface de la bactérie ou sur la surface d'un bactériophage dérivé de la bactérie, et l'analyse du marqueur dans un format à haut débit. L'invention concerne également un anticorps glycosylé comprenant une portion Fv qui reconnaît un antigène natif et s'y lie et une portion Fc qui est glycosylée sur un résidu asparagine conservé.

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