Application of fucosylation inhibitors for production of afucosylated antibody

Abstract Fucosylation is an important quality attribute for therapeutic antibodies. Afucosylated antibodies exhibit higher therapeutic efficacies than their fucosylated counterparts through antibody‐dependent cellular cytotoxicity (ADCC) mechanism. Since higher potency is beneficial in reducing dose or duration of the treatment, afucosylated antibodies have attracted a great deal of interest in biotherapeutics development. In this study, novel small molecules GDP‐D‐Rhamnose and its derivatives (Ac‐GDP‐D‐Rhamnose and rhamnose sodium phosphate) were synthesized to inhibit the enzyme in the GDP‐fucose synthesis pathway. Addition of these compounds into cell culture increased antibody afucosylation levels in a dose‐dependent manner and had no significant impact on other protein quality attributes. A novel and effective mechanism to generate afucosylated antibody is demonstrated for biologics discovery, analytical method development, process development, and other applications.