Metabolic engineering of monoclonal antibody carbohydrates for antibody-drug conjugation.
The role that carbohydrates play in antibody function and pharmacokinetics has made them important targets for modification. The terminal fucose of the N-linked glycan structure, which has been shown to be involved in modulation of antibody-directed cellular cytotoxicity, is a particularly interesting location for potential modification through incorporation of alternative sugar structures. A library of fucose analogues was evaluated for their ability to incorporate into antibody carbohydrates in place of the native fucose. A number of efficiently incorporated molecules were identified, demonstrating the ability of fucosyltransferase VIII to utilize a variety of non-natural sugars as substrates. Among these structures was a thiolated analogue, 6-thiofucose, which was incorporated into the antibody carbohydrate with good efficiency. This unnatural thio-sugar could then be used for conjugation using maleimide chemistry to produce antibody-drug conjugates with pronounced cytotoxic activities and improved homogeneity compared to drug attachment through hinge disulfides.
Seattle Genetics, Inc. , Bothell, Washington 98021, United States.
Nicole M
NM
Okeley
Brian E
BE
Toki
Xinqun
X
Zhang
Scott C
SC
Jeffrey
Patrick J
PJ
Burke
Stephen C
SC
Alley
Peter D
PD
Senter
D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000920:Antibody-Dependent Cell Cytotoxicity; D002241:Carbohydrates / Q000737:chemistry* / Q000276:immunology; D002460:Cell Line; D004220:Disulfides / Q000737:chemistry; D005643:Fucose / Q000031:analogs & derivatives* / Q000276:immunology; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry* / Q000276:immunology; D060847:Metabolic Engineering
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D004220:Disulfides; D018796:Immunoconjugates; D005643:Fucose