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TransGLYCIT® – Transglycosylation of IgG
TransGLYCIT is a platform technology that enables efficient and site-specific human IgG glycan remodeling. With TransGLYCIT, antibodies with defined and homogenous glycoforms are generated using fast and robust enzymatic workflows.
TransGLYCIT Remodeling generates a homogenous pool of antibodies carrying the G0, G1, G2 or G2S2 glycan profile with the core fucosylation intact, and TransGLYCIT Remodeling Afucosylated generates a homogenous pool of antibodies carrying the G0, G1, G2, G2S2 or Man5 glycan profile with core afucosylation.
Why TransGLYCIT®?
- Efficient and site-specific human IgG Fc N-glycan remodeling
- Prepares human IgG with homogenous glycoprofiles
- A rapid workflow – transglycosylation is completed within a few hours
Human IgG
3 hour workflow
G0, G1, G2, G2S2 or Man5 glycoform is included
Transglycosylation occurs at the Fc N-glycan sites
Product Formats
TransGLYCIT® Remodeling
Glycan remodeling of human IgG with the G0, G1, G2 or G2S2 glycoform
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TransGLYCIT® Remodeling Afucosylated
Glycan remodeling of human IgG with the G0, G1, G2, G2S2 or Man5 glycoform, with core afucosylation.
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About TransGLYCIT®
How TransGLYCIT works
- Deglycosylation
The Fc N-glycans are trimmed to the core GlcNAc using the IgG-specific GlycINATOR™ Immobilized (EndoS2) enzyme that hydrolyses all Fc glycoforms, including high-mannose, hybrid, complex and bisecting glycans. For TransGLYCIT Remodeling, the degree of core fucosylation is the same as on the original IgG. It is possible to obtain afucosylated G0, G1, G2, G2S2 or Man5 glycoforms using TransGLYCIT Remodeling Afucosylated, where the FucosEXO® 16 Immobilized 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 Man5 glycoform and the core GlcNAc.
The transglycosylation of IgG is followed by an affinity purification step.
The enzymes in TransGLYCIT are expressed in E. coli and modified with His-tags.
Applications
Functional comparison of antibody glycoforms by combining glycoengineering with ADCC and ADCP reporter assays to assess Fc effector activity.
Performance Activity
Simple and rapid generation of IgG with homogeneous G0F or G0 glycoforms for structural and functional characterization.
Rapid generation of galactosylated IgG with or without core fucose for structural and functional characterization.
Rapid and simple generation of IgG with defined Man5 glycoforms for structural and functional characterization.
Rapid generation of sialylated IgG Fc-glycan profiles, with or without core fucose, for structural and functional characterization.
Citations
Resources
TransGLYCIT® to Evaluate Fc Effector Function of Therapeutic Antibodies
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TransGLYCIT® for Fc-specific N-glycan Remodeling of Human IgG
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SmartStories
TransGLYCIT™ in a Study on Antibody Characterization
Guusje van Schaick
Leiden University Medical Center
FAQ and Support
Does TransGLYCIT work on any glycoprotein?
No, TransGLYCIT is based on IgG specific enzymes and will only transglycosylate 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.
What antibody species can I transglycosylate using TransGLYCIT?
The TransGLYCIT platform is developed for transglycosylation of human IgG.
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.
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.
Legal & Disclaimers
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Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals. All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.
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