Generation of IgG with Galactosylated Glycan Profiles using TransGLYCIT®

Performance Activity

Rapid generation of galactosylated IgG with or without core fucose for structural and functional characterization.

Terminal galactosylation of the IgG Fc glycans result in monogalactosylated (G1) or digalacosylated (G2) glycoforms, with additional fucosylated glycoform variants (G1F and G2F). By generating defined G1 and G2 glycan profiles, the differential impact of galactosylation on the structural and functional properties of the antibody can be analyzed and compared with other defined glycoforms, including both fucosylated and afucosylated glycan profiles.

Galactosylated glycan profiles with and without core fucose were obtained by transglycosylation of trastuzumab using the TransGLYCIT Remodeling and TransGLYCIT Remodeling Afucosylated kits with G1 or G2 glycoforms. Analysis at subunit level using LC-MS (Fig. 1, 2) show mass spectra of the native glycan profile of trastuzumab and the mass shifts after transglycosylation to generate G1F and G2F glycoforms, or afucosylated G1 or G2 glycoforms.

Simple generation of galactosylated IgG with or without core fucose

Figure 1. Generation of IgG with homogeneous G1 or G1F glycoforms. Deconvoluted mass spectra of the scFc fragment of native trastuzumab (top) and after transglycosylation with TransGLYCIT Remodeling G1 (middle), or TransGLYCIT Remodeling Afucosylated G1 (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).
Figure 2. Generation of IgG with homogeneous G2 or G2F glycoforms. Deconvoluted mass spectra of the scFc fragment of native trastuzumab (top) and after transglycosylation with TransGLYCIT Remodeling G2 (middle), or TransGLYCIT Remodeling Afucosylated G2 (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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