Transglycosylation of IgG using TransGLYCIT
TransGLYCIT is a platform that enables efficient and site-specific human IgG glycan remodeling and conjugation. With TransGLYCIT, antibodies with defined and homogenous glycoforms or site-specific azide-activation are generated using fast and robust enzymatic workflows.
With the optional FucosEXO™ 16 enzyme in the glycan remodeling workflow, afucosylated antibodies carrying the Fc glycoform of choice (G0, G1, G2 or G2S2) can be obtained for comparison of antibodies with or without core fucose. With the azide activation workflow, antibodies are prepared for conjugation with any label of choice using click-chemistry.
TransGLYCIT is available in the following product formats:
- TransGLYCIT (glycan remodeling)
Generation of 1 mg human IgG with the G0, G1, G2 or G2S2 glycoform. - TransGLYCIT Afucosylated (glycan remodeling)
Generation of 1 mg afucosylated human IgG with the G0, G1, G2 or G2S2 glycoform. - TransGLYCIT Azide Activation NEW!
Azide activation of 100 µg human IgG1 or IgG4. - TransGLYCIT Azide Activation hIgG2 NEW!
Azide activation of 100 µg human IgG2.
The Transglycosylation Workflow
- 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. The degree of fucosylation is the same as on the original IgG. Note that for the glycan remodeling products, it is possible to obtain afucosylated G0, G1, G2 or G2S2 glycoforms using TransGLYCIT Afucosylated, where the immobilized FucosEXO 16 enzyme hydrolyzes the α1,6-linked core fucose prior to the transglycosylation step. - Transglycosylation
The engineered glycosynthase TransINATOR™ catalyzes the transglycosylation reaction between the oxazoline-reactive G0, G1, G2, G2S2 or azide-activated glycoform and the core GlcNAc.
The transglycosylation of IgG is followed by an affinity purification step.
About TransGLYCIT
TransGLYCIT is a platform technology for efficient transglycosylation of native human IgG in a few hours. The robust workflow transglycosylates the antibody Fc N-glycans using enzymatic remodeling, resulting in a homogenous pool of antibodies carrying a defined glycoform. The technology is available in kit formats to generate antibodies carrying the G0, G1, G2 or G2S2 glycan profile, or to generate azide-activated antibodies carrying an azide-activated glycoform. With the TransGLYCIT Afucosylated kit, antibodies without the core fucose are generated. The enzymes in TransGLYCIT are expressed in E. coli and modified with His-tags. TransGLYCIT is provided in kits for processing of 100 µg (azide activation kits) or 1 mg (glycan remodeling kits) of human IgG.
For content and storage information, please see the product pages.
- Human IgG1, IgG2 and IgG4
- 3-6 hours workflow
- G0, G1, G2, G2S2 or azide-activated glycoform is included
- Transglycosylation occurs at the Fc N-glycan sites
Applications
Product Documents
Certificate of Analysis
FAQ
Popular FAQ
GlycINATOR is an IgG specific endoglycosidase that hydrolyzes complex, hybrid and high mannonse type glycans on the conserved Fc site on IgG.
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.
Unfortunately no, the enzyme requires trimming of the Fc-glycan using GlycINATOR to enable access to the core fucose substrate.
No, TransGLYCIT is based on IgG specific enzymes and will only transglycosylate IgG.
The TransGLYCIT platform is developed for transglycosylation of human IgG.
