Monoclonal antibodies (mAbs) are an important class of therapeutic proteins widely used for the treatment of cancer, inflammation, and infectious diseases. It has been demonstrated that the presence and nature of Fc domain glycosylation can profoundly affect the effector functions of antibodies, including the antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity, and anti-inflammatory activity. Therefore, controlling glycosylation of a therapeutic monoclonal antibody to a desired glycosylation status is important for its therapeutic efficacy. Nevertheless, most of monoclonal antibodies on the market are produced as mixtures of glycoforms that are not optimal for therapeutic applications, due to the difficulties in controlling protein glycosylation to a defined homogeneous status. This presentation describes an enzyme-based glycosylation remodeling method for site-specific glycoengineering of antibody’s Fc and Fab glycans in an intact antibody. We have found that a remarkable site-selectivity in glycosylation remodeling can be achieved by taking advantage of the substrate specificity of a handful of endoglycosidases and glycosidases and their mutants. The combined use of the enzyme specificity permits site-selective introduction of different glycans at different sites (Fab and Fc) in a given antibody, which is hitherto difficult to achieve by glycoengineering at genetic level. Examples for glycoengineering of several therapeutic monoclonal antibodies for improving their effector functions will be provided. The use of the chemoenzymatic method for making antibody-drug conjugates (ADCs) will be also discussed.

Chemoenzymatic glycoengineering of antibodies: Enzyme substrate specificity is the name of the game

251st National Meeting (2016)

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