Glycosylation profile analysis

C - Chemistry – Metallurgy – 12 – P

Patent

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Details

C12P 21/00 (2006.01)

Patent

CA 2697091

The present invention provides a method for the production of a glycosylated heterologous polypeptide comprising the steps of obtaining a sample from a crude fermentation broth, incubation of the sample with magnetic affinity beads, releasing glycans from the immobilized glycosylated polypeptides, measuring a glycosylation profile, comparing the glycosylation profile with a desired glycosylation profile of the recombinant glycosylated polypeptide, modifying the culture conditions in accordance to the glycosylation profile obtained, and repeating the process in order to obtain a glycosylated heterologous polypeptide with the desired glycosylation profile. With a similar method diagnostic markers can be identified and quantified.

La présente invention concerne un procédé de production d'un polypeptide hétérologue glycosylé comprenant les étapes consistant à obtenir un échantillon d'un bouillon de fermentation brut, à incuber l'échantillon avec des billes magnétiques pour chromatographie d'affinité, à libérer les glycanes des polypeptides glycosylés immobilisés, à mesurer un profil de glycosylation, à comparer le profil de glycosylation avec un profil de glycosylation désiré pour le polypeptide glycosylé recombinant, à modifier les conditions de culture en fonction du profil de glycosylation obtenu et à répéter le processus afin d'obtenir un polypeptide hétérologue glycosylé présentant le profil de glycosylation désiré. Des marqueurs diagnostiques peuvent être identifiés et quantifiés par un procédé similaire.

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