Powered by BiozGlyCLICK®
Download Product Folder
GlyCLICK® – Site-specific Conjugation of IgG
GlyCLICK is a three-step conjugation technology for site-specific and quantitative conjugation of IgG from several species and subclasses.
With GlyCLICK, specific Fc N-glycan hydrolysis allows for site-specific conjugation at the core GlcNAcs using robust click-chemistry and results in a degree of label (DOL) or drug-antibody ratio (DAR) of 2. The reliable performance ensures quantitative conjugation and intact immunoreactivity for sensitive applications.
The technology is available in a range of kit formats, including conjugation with fluorophores, biotin, DFO, or toxins for ADC development. The format “Azide Activation” renders the antibody azide-activated, for conjugation with any alkyne-reactive label of choice.
Why GlyCLICK®?
- Site-specific conjugation of IgG
- Reliable performance ensuring quantitative conjugation and preserved immunoreactivity for sensitive applications
- Stable and homogenous conjugation generates a degree of labeling (DOL) of 2
Human IgG1-4, IgG from mouse, rabbit, rat, monkey, sheep, goat, cow and horse
2-3 day protocol
Fluorophore, Biotin, DFO, MMAE, PNU, Azide Activation
Conjugation occurs at the Fc N-glycan sites
Product Formats
GlyCLICK® Azide Activation
Site-specific conjugation of IgG with azide-alkyne click chemistry
Buy product
About GlyCLICK®
How GlyCLICK works
- Deglycosylation
The Fc specific endoglycosidase GlycINATOR® (EndoS2) hydrolyzes the Fc glycans to the inner most GlcNAc moiety on several subclasses and species of IgG. The enzyme removes all glycoforms, including; high-mannose, hybrid, complex, and bisecting type glycans. - Azide Activation
Azide-containing UDP-GalNAz is enzymatically attached to the exposed GlcNAc using the ß-1,4-Galactosyltransferase Y289L* (GalT) to generate an azide-activated antibody that is reactive with an alkyne-carrying label. - CLICK Reaction
The azide activated antibody is conjugated with a cyclooctyne-functionalized label of choice in a biorthogonal reaction through strain-promoted copper-free click chemistry (SPAAC) to form a stable triazole.
Applications
Site-specific ADC generation with defined DAR, preserving antigen binding and delivering potent, controlled cytotoxicity in target cells.
Improved PET/CT tumor targeting and biodistribution using site-specific antibody labeling with consistent DOL and preserved in vivo performance.
Supports high-quality immunofluorescence imaging with consistent signal intensity and reliable quantification through defined antibody labeling.
Higher-resolution flow cytometry with improved signal intensity and a 10-fold increase in separation index using site-specific antibody labeling.
Citations
Resources
GlyCLICK® for Generation of Dual-payload and Bispecific ADCs
Download Scientific Poster
GlyCLICK® for Generation of Improved ADCs
Download Scientific Poster
GlyCLICK® for Dual-payload ADCs
Download Scientific Poster
GlyCLICK®: From Kit to Clinic
Watch video
GlyCLICK® for Generation and Analysis of ADCs
Download Scientific Poster
GlyCLICK® for Generation of Site-specific Conjugates for Fluorescent Imaging
Download Scientific Poster
FAQ and Support
Which isotopes can I use to radiolabel IgG after chelation using GlyCLICK?
The GlyCLICK technology can be used to conjugate IgG with Desferrioxamine (DFO) using the GlyCLICK DFO kit. The DFO is a chelating agent for radiolabeling with the radioisotope Zirconium-89 (89Zr) for PET-imaging. The Azide Activation kit can be used to conjugate a chelator of choice if it is alkyne-modified (carrying for example DIBO, DBCO or BCN) to be compatible with the click chemistry.
Why do I need to use an alkyne-functionalized label with GlyCLICK?
The final step of GlyCLICK labeling kit is based on a copper free click reaction (SPAAC) and it is a strain promoted reaction between an azide and an alkyne-carrying label. As the name indicates the reaction occurs spontaneously due to the strained bond angles. The use of this click reaction that forms stable conjugates, allows you to freely choose label or payload to combine with your IgG as long as it is alkyne-functionalized. Several alkyne-carrying labels are commercially available with cyclooctyne structures such as DIBO, DBCO or BCN.
What is the conjugation efficacy using GlyCLICK?
All the steps (deglycosylation, azide activation and click reaction) in GlyCLICK are highly efficient. Normally, all molecules are conjugated when using GlyCLICK, resulting in DOL=2. On the application page we have shown by MS-analysis that all molecules have been conjugated and there is no remaining un-conjugated material.
What fluorescent labels can I use with the GlyCLICK technology?
GlyCLICK is available in kit formats with AlexaFluor®488, AlexaFluor®555, AlexaFluor®647 or in the Azide Activation kit format for conjugation of a label of choice. With the fluorescent label being alkyne-modified (carrying for example DIBO, DBCO or BCN) it is possible to combine it with the GlyCLICK azide activation kit.
What is the smallest amount of IgG that I can process using the 250 ug kit?
The GlyCLICK 250 ug kit is optimized for 250 ug of IgG as a starting material. We do not recommend using less than 100 ug of starting material in a minimum of 100µL to achieve proper end-over-end mixing on the GlycINATOR columns. Please contact support@genovis.com for assistance if you want to process smaller amounts then 100µg.
Legal & Disclaimers
All rights reserved. Genovis products may be covered by one or more patents, trademarks and copyrights owned by Genovis AB or licensed from third parties. For more information about commercial rights, please contact the Genovis team at licensing@genovis.com.
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.
© Genovis AB