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TransGLYCIT Generates Homogenous IgG Glycoforms

TransGLYCIT is a platform that enables specific IgG glycan remodeling and prepares antibodies with defined and homogenous glycoforms 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 core fucose.

TransGLYCIT is available in the following product formats:

TransGLYCIT 1 mg
Generation of up to 1 mg human IgG with the G0, G1, G2 or G2S2 glycoform.
TransGLYCIT 1 mg Afucosylated
Generation of up to 1 mg afucosylated human IgG with the G0, G1, G2 or G2S2 glycoform.

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. To obtain afucosylated glycoforms, the optional Immobilized FucosEXO™ 16 enzyme hydrolyzes the α1-6 linked core fucose.
Transglycosylation
The engineered glycosynthase TransINATOR™ catalyzes the transglycosylation reaction between the oxazoline reactive G0, G1, G2 or G2S2 glycoform and the core GlcNAc.

TransGLYCIT Product Information

TransGLYCIT is a platform technology for efficient transglycosylation of native IgG in three hours. The robust workflow transglycosylates the antibody Fc N-glycans using enzymatic remodeling, resulting in a homogenous pool of antibodies with a defined glycoform. The technology is available in kit formats to generate antibodies carrying the G0, G1, G2 or G2S2 glycan profile with the option to remove the core fucose using the TransGLYCIT Afucosylated kit. The enzymes in TransGLYCIT are expressed in E. coli and modified with His-tags. TransGLYCIT is provided in kits for processing of up to 1 mg of human IgG.

Unit Definition

TransGLYCIT transglycosylates ≥ 95% human IgG1 as measured by LC-MS.

Storage and Content

TransGLYCIT is shipped cold, and the content should be stored at different temperatures upon arrival. Do not freeze the product! TransGLYCIT is provided in kits containing sufficient material to transglycosylate up to 1 mg of IgG. TransGLYCIT is for R&D use only.

TransGLYCIT contains:

Immobilized GlycINATOR (TransGLYCIT)
Immobilized GlycINATOR & Immobilized FucosEXO 16 (TransGLYCIT Afucosylated)
TransINATOR enzyme
Oxazoline glycoform G0, G1, G2 or G2S2
CaptureSelect™ spin column for purification

FAQ

Popular FAQ

What is the substrate specificity of GlycINATOR?

GlycINATOR is an IgG specific endoglycosidase that hydrolyzes complex, hybrid and high mannonse type glycans on the conserved Fc site on IgG.

Does TransGLYCIT work on all human IgG subclasses?

Yes, the transglycosylation reaction can be performed on all subclasses of human IgG. The reaction is somewhat slower on IgG2 and longer incubation times may be necessary to obtain over 95% transglycosylation.

Can I use the FucosEXO 16 enzyme on native antibodies to remove the core fucose?

Unfortunately no, the enzyme requires trimming of the Fc-glycan using GlycINATOR to enable access to the core fucose substrate.

Does TransGLYCIT work on any glycoprotein?

No, TransGLYCIT is based on IgG specific enzymes and will only transglycosylate IgG.

What antibody species can I transglycosylate using TransGLYCIT?

The TransGLYCIT platform is developed for transglycosylation of human IgG.

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TransGLYCIT Generates Homogenous IgG Glycoforms

The Transglycosylation Workflow