METHOD FOR MODULATING AFUCOSLYATION OF AN ANTIBODY PRODUCT
The present application is directed to methods for modulating afucosylation of an antibody product that is produced in a bioreactor. The methods include the addition of mannose to the bioreactor to control the afucosylation of the antibody product, including increasing by afucosylation by 1% or more, relative to untreated bioreactor products.
1 . A method for increasing afucosylation of an antibody recombinantly expressed in a bioreactor, the method comprising:
culturing cells that express the antibody in a bioreactor;
adding mannose to cell culture during the antibody production process such that the afucosylation of the antibody is increased as compared to the same antibody produced by the antibody production process without the addition of mannose.
2 . The method of claim 1 , wherein the amount of mannose added is from about 1 g/L to about 10 g/L.
3 . The method of claim 1 , wherein the bioreactor has a volume of at least 10 L.
4 . The method of claim 1 , wherein the antibody is a monoclonal IgG.
5 . The method of claim 1 , wherein the cells are mammalian.
6 . The method of claim 1 , wherein afucosylation of the antibody, is increased by at least 0.5% as compared to the antibody produced by the antibody production process without the addition of mannose.
7 . The method of claim 6 , wherein afucosylation of the antibody product is increased by at least 1%, such as at least 2%, as compared to the antibody produced by the antibody production process without the addition of mannose.
8 . The method of claim 1 , wherein the antibody production process is a fed-batch process.
9 . The method of claim 1 , wherein the antibody production process is a perfusion process.
10 . The method of claim 1 , which further comprises isolating the expressed antibody and optionally subjecting the antibody to one or more purification steps.
11 . A method for matching the afucosylation of a recombinantly produced antibody to a previously obtained target afucosylation percentage for the same antibody, the method comprising:
culturing cells that express the antibody in a bioreactor;
controlling the addition of mannose to the bioreactor during the antibody production process to obtain the expressed antibody with the target percentage afucosylation.
12 . The method according to claim 11 wherein the percentage afucosylation of the expressed antibody is within +/−0.25% of the target percentage afucosylation.
13 . The method of claim 11 , wherein the antibody production process is a perfusion process.