Hoffmann-La Roche Inc.

Roche Diagnostics Gmbh

F. Hoffmann-La Roche Ag

Heiko Walch

Harald Sobek

Roberto Falkenstein

Marco Thomann

Sebastian Malik

Matthias Freiherr Von Roman

Ingrid Grunert

Thomas Dams

Ingrid Hoffmann

Roland Dorn

Michael Hingar

Kiyoshi Yasukawa

Christian Helmut Bell

Misato Baba

Herein is reported a method for the enzymatic preparation/production of an antibody with a modified glycosylation in the Fc-region comprises the steps of incubating an antibody that has a glycosylation in the Fc-region with one or more enzymes for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region to a defined form, separating in one or more chromatography steps the antibody with a modified glycosylation in the Fc-region from the one or more enzymes and thereby producing the antibody with a modified glycosylation in the Fc-region and one or more recycled enzymes, and repeating the incubating step with the one or more recycled enzymes at least once.

Re-use of enzymes in in vitro glycoengineering of antibodies

Herein is reported a method for the enzymatic production of an antibody with a modified glycosylation in the Fc-region comprising the steps of incubating the antibody light chain affinity ligand-bound monoclonal antibody with a glycosylation in the Fc-region with a first enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region, recovering the antibody from the antibody light chain affinity ligand, incubating the recovered antibody in solution with a second enzyme for a time sufficient and under conditions suitable to modify the glycosylation of the Fc-region to a defined form, separating the antibody with the modified glycosylation in the Fc-region from the second enzyme in a cation exchange chromatography, and thereby producing the antibody with a modified glycosylation in the Fc-region.

Method for in vitro glycoengineering of antibodies

Growing community of inventors

Penzberg, Germany

Heiko Walch