G - Physics – 01 – N
Patent
G - Physics
01
N
G01N 33/68 (2006.01) C07K 1/00 (2006.01) C07K 1/14 (2006.01) C07K 1/16 (2006.01) C07K 1/22 (2006.01) C07K 16/06 (2006.01)
Patent
CA 2750977
The current invention is directed to a method for the determination of the glycosylation pattern of a human immunoglobulin of the subclass IgG1 or IgG4 or of a murine immunoglobulin of the subclass IgG2a or IgG3 comprising the following steps: a) cleaving said immunoglobulin into fragments by enzymatic digestion with the enzyme IdeS, b) separating the fragments of said immunoglobulin obtained by the enzymatic digestion by reversed phase high performance liquid chromatography, c) subjecting the separated fragments of said immunoglobulin obtained in step b) to a mass spectrometric analysis, and d) determining the glycosylation pattern of said immunoglobulin from the mass spectrometric data obtained in step c).
La présente invention porte sur un procédé de détermination du motif de glycosylation d'une immunoglobuline humaine de la sous-classe IgG1 ou IgG4 ou d'une immunoglobuline murine de la sous-classe IgG2a ou IgG3 comprenant les étapes suivantes consistant à : a) cliver ladite immunoglobuline en fragments par digestion enzymatique par l'enzyme IdeS, b) séparer les fragments de ladite immunoglobuline obtenus par la digestion enzymatique par une chromatographie liquide haute performance en phase inverse, c) soumettre les fragments séparés de ladite immunoglobuline obtenus à l'étape b) à une analyse par spectrométrie de masse et d) déterminer le motif de glycosylation de ladite immunoglobuline à partir des données de spectrométrie de masse obtenues à l'étape c).
Koll Hans
Regula Joerg Thomas
Sondermann Peter
Zeck Anne
Gowling Lafleur Henderson Llp
Roche Glycart Ag
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