GlyCLICK™

GlyCLICK is a site-specific conjugation technology for antibodies based on enzymatic remodeling of the Fc glycans and click chemistry. The technology covalently conjugates a label of choice to the Fc domain of the antibody with a degree of label of 2.0, resulting in a homogenous antibody conjugate with preserved antigen binding.

GlyCLICK is available for conjugation of 250 μg of antibody with the following DIBO conjugates: Alexa Fluor® 488, biotin and the chelator DFO. The kit is also available with azide activation-only, for custom conjugation with a cyclooctyne label. This azide activation kit is available for conjugation of 1 x 250 μg and 1 x 10 mg of antibody.

Product Description

GlyCLICK is a unique conjugation technology for site-specific labeling of native antibodies at the N-linked glycans in the Fc domain. The IgG-specific endoglycosidase GlycINATOR (EndoS2) is utilized for its ability to hydrolyze IgG Fc glycans after the core GlcNAc¹. Using an engineered galactosyl transferase, GalT(Y289L), a GalNAz residue is subsequently attached to the exposed GlcNAc, resulting in an azide available for a copper-free biocompatible strain-promoted azide-alkyne click-chemistry reaction² with a cyclooctyne label³.

A wide variety of cyclooctyne labels, such as DIBO-labels (functional groups), can be specifically conjugated to the antibody including drugs, chelators, biotin or fluorophores. Using GlyCLICK, functional groups can be conjugated to the Fc domain of antibodies of several species and a variety of subclasses. The technology results in a ratio of 2 conjugates per antibody with high reproducibility and a low batch-to-batch variation. GlyCLICK is applicable to all IgG antibodies carrying Fc glycans and does not rely on prior engineering of the amino acid sequence.

The applications for the GlyCLICK-technology includes e.g. cytotoxic drug development, in vitro and in vivo cell imaging, immunohistochemistry and western blotting.

• Learn more about the applications of the GlyCLICK-technology

(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 α1-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.

How GlyCLICK works

IgG contain a conserved N-linked glycosylation site on the CH2 domain of each heavy chain of the Fc region. This site of the antibody is used by GlyCLICK for selective conjugation of a label of choice. The labeling is performed in three steps:

1. Immobilized GlycINATOR (EndoS2) hydrolyzes the N-glycans on the Fc domain of the IgG, leaving the innermost GlcNac residue intact.
2. Specific attachment of a GalNAz residue to the core GlcNAc by an engineered galactose transferase (GalT(Y298L)).
3. The azide-activated antibody reacts with a DIBO-alkyne label in a strain-promoted, copper-free click chemistry reaction and forms a stable and homogenous antibody conjugate.

Patent and Disclaimer

GlyCLICK™

GlycINATOR®

All rights reserved. Aspects of GlycINATOR technology are encompassed by pending patent applications and by granted Patent in Singapore, China, USA and EPO in the name of Genovis AB. The trademark GlycINATOR is the property of Genovis AB.

The trademark GlyCLICK is the property of Genovis AB.

For research use only. Not intended for any animal or human therapeutic or diagnostic use.

All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.

SiteClick™

This product is provided under an intellectual property license from Life Technologies Corporation.  The transfer of this product is conditioned on the buyer using the purchased product solely in research conducted by the buyer, excluding contract research or any fee for service research, and the buyer must not (1) use this product or its components for (a) diagnostic, therapeutic or prophylactic purposes; (b) testing, analysis or screening services, or information in return for compensation on a per-test basis; or (c) manufacturing or quality assurance or quality control, and/or  (2) sell or transfer this product or its components for resale, whether or not resold for use in research.  For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

The trademark SiteClick™ is the property of Life Technologies Corporation.

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