Generation of IgG with Homegeneous Man5 Glycan Profiles using TransGLYCIT®

Performance Activity

Rapid and simple generation of IgG with defined Man5 glycoforms for structural and functional characterization.

Of the high-mannose N-glycan structures, Man5 is the most common one present on therapeutic antibodies. Antibodies carrying Man5 have shown increased clearance rates compared to those carrying complex N-glycan structures. With TransGLYCIT, defined Man5 glycoforms of therapeutic antibodies can be generated to investigate the impact of this N-glycan structure on functional antibody properties such as serum half-life and clearance.

High-mannose N-glycan structures are biosynthetically generated without core fucose. Using the TransGLYCIT platform, trastuzumab was remodeled to carry the Man5 glycoform within three hours. The transglycosylated antibody was analyzed at the subunit level using LC-MS. The mass spectra show the glycan profile of native trastuzumab, and the resulting homogeneity and mass shift after complete transglycosylation of the antibody using TransGLYCIT Remodeling Afucosylated Man5.

Simple generation of IgG with homogeneous Man5 glycans

Generation of IgG with homogeneous Man5 glycoforms. Deconvoluted mass spectra of the scFc fragment of native trastuzumab (top) and after transglycosylation with TransGLYCIT Remodeling Afucosylated Man5 (bottom). The mAb was digested with FabRICATOR and the subunits analyzed by reversed-phase LC-MS on a Waters™ BioAccord™ system equipped with a Waters™ BioResolve RP mAb column (2.1 x 50 mm).

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