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GlyCLICK^®

GlyCLICK is a site-specific conjugation technology for IgG using Fc glycan remodeling and click-chemistry.

The technology generates stable and homogenous antibody conjugates for IgG from several species and subclasses. Fc-glycan remodeling by complete deglycosylation of the antibody allows for site-specific conjugation using robust click-chemistry, resulting in a degree of label (DOL) or antibody-drug ratio (DAR) of 2.0.

The reliable performance ensures quantitative conjugation and intact immunoreactivity for sensitive applications. The technology is available in a range of kit formats to facilitate tailored conjugates by site-specific labeling of antibodies using customized labels.

Alexa Fluor 488

In vitro cell imaging, IHC, FCM

Alexa Fluor 555

In vitro cell imaging, IHC, FCM

Alexa Fluor 647

In vitro cell imaging, IHC, FCM

Biotin

Immunoassays, ELISA, WB

DFO

Immuno- and In vivo Imaging

ADC MMAE

ADC Development

ADC PNU

ADC Development

Azide Activation

Custom Conjugation

How GlyCLICK works

Deglycosylation
The Fc specific Endoglycosidase GlycINATOR^®(EndoS2)¹hydrolyzes 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 UPD-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 functionalized label²of choice in a biorthogonal reaction through strain-promoted copper-free click chemistry (SPAAC)³ to form a stable triazole.

Characteristics

GlyCLICK enables site-specific and quantitative conjugation of IgG from several species and subclasses with a degree of label (DOL) of 2. The specificity of GlycINATOR allows for the complete deglycosylation of all IgG Fc-glycoforms and does not require any prior engineering of the amino acid sequence. Robust conjugation using click-chemistry results in stable and completely labeled antibody conjugates with minimized the variation between batches.

Contact

For custom made formats or licensing opportunities of GlyCLICK, please contact the Genovis team.

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References

1. Sjögren, J. et al., 2013. EndoS2 is a unique and conserved enzyme of serotype M49 group A Streptococcus that hydrolyses N-linked glycans on IgG and a1-acid glycoprotein. The Biochemical Journal, 455(1), pp.107–118.
2. SiteClick™ is provided under an intellectual property license from Life Technologies Corporation. The trademark SiteClick™ is the property of Life Technologies Corporation.
3. Ramakrishnan, B. & Qasba, P.K., 2002. Structure-based design of beta 1,4-galactosyltransferase I (beta 4Gal-T1) with equally efficient N-acetylgalactosaminyltransferase activity: point mutation broadens beta 4Gal-T1 donor specificity. J Biol Chem, 277(23), pp.20833–20839.

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