Beta-Carbafucose inhibits cellular fucosylation to enable efficient production of afucosylated antibodies
A paper detailing our foundational research is now available at PNAS. Click below to access the paper, or read the abstract below.
Abstract
The fucosylation of glycoproteins regulates diverse physiological processes. Inhibitors that can control cellular levels protein fucosylation have consequently emerged as being of high interest. One area where inhibitors of fucosylation have gained significant attention is for the production of afucosylated antibodies, which exhibit superior antibody-dependent cell cytotoxicity (ADCC) as compared to their fucosylated counterparts. Here we describe b-carbafucose, a fucose derivative in which the endocyclic ring oxygen is replaced by a methylene group, and show that it acts as a potent metabolic inhibitor within cells to antagonize protein fucosylation. b-Carbafucose is assimilated by the fucose salvage pathway to form GDP-carbafucose which, due to its being unable to form the oxocarbenium ion-like transition states used by fucosyltransferases, is an incompetent substate for these enzymes. Treating a CHO cell line used for high-level production of the therapeutic antibody Herceptin leads to dose dependent reductions in core fucosylation without affecting cell growth or antibody production. Mass spectrometry analyses of the intact antibody and N-glycans, shows that b-carbafucose is not incorporated into the antibody N-glycans at detectable levels. We expect that b-carbafucose will serve as a useful research tool for the community and may find immediate application for the rapid production of afucosylated antibodies for therapeutic purposes.