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Optimal Synthetic Glycosylation of a Therapeutic Antibody
Author(s) -
Parsons Thomas B.,
Struwe Weston B.,
Gault Joseph,
Yamamoto Keisuke,
Taylor Thomas A.,
Raj Ritu,
Wals Kim,
Mohammed Shabaz,
Robinson Carol V.,
Benesch Justin L. P.,
Davis Benjamin G.
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201508723
Subject(s) - glycosylation , antibody , glycation , chemistry , endoglycosidase , computational biology , combinatorial chemistry , biochemistry , fragment crystallizable region , biology , immunology , receptor
Glycosylation patterns in antibodies critically determine biological and physical properties but their precise control is a significant challenge in biology and biotechnology. We describe herein the optimization of an endoglycosidase‐catalyzed glycosylation of the best‐selling biotherapeutic Herceptin, an anti‐HER2 antibody. Precise MS analysis of the intact four‐chain Ab heteromultimer reveals nonspecific, non‐enzymatic reactions (glycation), which are not detected under standard denaturing conditions. This competing reaction, which has hitherto been underestimated as a source of side products, can now be minimized. Optimization allowed access to the purest natural form of Herceptin to date (≥90 %). Moreover, through the use of a small library of sugars containing non‐natural functional groups, Ab variants containing defined numbers of selectively addressable chemical tags (reaction handles at Sia C1) in specific positions (for attachment of cargo molecules or “glycorandomization”) were readily generated.