NEW TransGLYCIT™ Glycoforms Available!

Genovis launches new glycoforms with TransGLYCIT™ – A platform to create IgG with defined glycan profiles!

TransGLYCIT enables specific IgG N-glycan remodeling and prepares antibodies with a defined and homogenous G0, G1, G2 or G2S2 glycoform using fast and robust enzymatic workflows. With the optional FucosEXO™ 16 enzyme, afucosylated antibodies can be obtained that enable direct comparison of antibodies with or without the core fucose.

1. Deglycosylation: The Fc N-glycans are trimmed to the core GlcNAc using the IgG specific Immobilized GlycINATOR® (EndoS2) enzyme that hydrolyses all Fc glycoforms including high-mannose, hybrid, complex and bisecting glycans. With the TransGLYCIT Afucosylated kits, Immobilized FucosEXO 16 hydrolyzes the α1-6 linked core fucose to obtain afucosylated glycoforms.
2. Transglycosylation: The engineered glycosynthase TransINATOR™ catalyzes the transglycosylation reaction between the exposed core GlcNAc and the oxazoline reactive G0, G1, G2 or G2S2 glycoform.

Generation of Defined Glycan Profiles using TransGLYCIT

Glycoengineering of the IgG glycan profile is important for the development of next-generation therapeutic antibodies with enhanced or silenced Fc effector functions. The generation of homogenous preparations is however challenging and often requires a complex bioprocess development or time-consuming cell-engineering strategies. The new TransGLYCIT platform remodels the IgG Fc N-glycans within three hours, resulting in antibodies carrying defined G0, G1, G2 or G2S2 glycan profiles, with or without the core fucosylation.

To demonstrate the transglycosylation workflow, the TransGLYCIT or TransGLYCIT Afucosylated kits were used to remodel the glycan profile of trastuzumab and the antibody was then analyzed by LC-MS at the subunit level (Fig. 1, 2). The resulting mass spectra show the heterogenous glycan profile of native trastuzumab (top) initially carrying both afucosylated and fucosylated Fc glycans. The mass shifts indicate a complete transglycosylation with the G1 or G2 glycoforms (middle) using the TransGLYCIT kit with the G1 or G2 glycoform. Homogenous and afucosylated glycan profiles carrying G1 or G2 glycoforms were also generated using the TransGLYCIT Afucosylated kit with the G1 or G2 glycoforms (bottom).

Figure 1, 2. Deconvoluted mass spectra of the Fc/2 fragment of native trastuzumab (top) and after transglycosylation with TransGLYCIT G1 or G2 (middle), or TransGLYCIT G1 or G2 Afucosylated (bottom). The mAb was digested with FabRICATOR® and the subunits were analyzed by reverse-phase LC-MS on a Waters BioAccord system equipped with a Waters BioResolve RP mAb column (2.1 x 50 mm).

TransGLYCIT 1 mg – Transglycosylation of up to 1 mg human IgG with the G0, G1, G2 or G2S2 glycoform.

TransGLYCIT 1 mg Afucosylated – Transglycosylation and afucosylation of up to 1 mg human IgG with the G0, G1, G2 or G2S2 glycoform.