Generation of IgG with G0 Glycan Profiles using TransGLYCIT®

Performance Activity

Simple and rapid generation of IgG with homogeneous G0F or G0 glycoforms for structural and functional characterization.

Agalactosylated IgG Fc glycans include both the fucosylated (G0F) and afucosylated (G0) glycoforms, where G0F accounts for a significant portion of the glycoforms observed in human serum and among several therapeutic antibodies. With TransGLYCIT, defined G0F and G0 glycan profiles can be generated for applications in both structural and functional analytical workflows for therapeutic antibodies. 

Using the TransGLYCIT platform, the glycan profile of a therapeutic antibody was remodeled to carry G0 glycoforms within three hours and analyzed at the subunit level using LC-MS. The resulting mass spectra show the heterogenous glycan profile of native trastuzumab, and the mass shifts after complete transglycosylation of the antibody to generate G0 glycoforms using TransGLYCIT Remodeling G0. A homogenous afucosylated glycan profile was generated using the TransGLYCIT Remodeling Afucosylated G0.

Efficient generation of IgG with homogeneous G0 or G0F glycoforms

Generation of homogeneous G0 glycoforms using TransGLYCIT. Deconvoluted mass spectra of the scFc fragment of native trastuzumab (top) and after transglycosylation with TransGLYCIT G0 (middle), or TransGLYCIT G0 Afucosylated (bottom). The mAb was digested with FabRICATOR and the subunits analyzed by reverse 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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