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 Formats
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.
(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.
Key Characteristics
- Human IgG1-4, Fc-fusion proteins, IgG from mouse, rabbit, rat, monkey, sheep, goat, cow and horse
- ~3 day protocol
- Available conjugates: AlexaFluor®488, biotin and DFO. Azide activation kits are available for custom conjugation
Applications
- ADC Development
Conjugate drugs to the Fc portion of an antibody using GlyCLICK - Fluorescent imaging
Efficient labeling of antibodies using GlyCLICK for fluorescent imaging - In vivo imaging
An antibody carrying a DFO chelator allows in vivo imaging - Intact Antigen Binding
Specific Fc conjugation ensures intact immunoreactivity - Specific labeling at the Fc
Conjugation of 2 labels per antibody
Unit Definition
GlyCLICK contains 250 units for conjugation of 250 μg of antibody. The azide activation kit is also available for conjugation of 10 mg of antibody. For larger amounts or for evaluation of GlyCLICK for ADC projects, please contact us using the contact form below.
GlyCLICK Contains
- Immobilized GlycINATOR
- UDP-GalNAz
- GalT(Y298L)
- DIBO label of choice
- All buffers needed
- Concentration and desalting columns
Download GlyCLICK Posters
Antibody Engineering 2017
"Site-specific Conjugation of Antibodies using GlyCLICK™ Results in Homogenous Conjugates, Preserved Antigen Binding, and Increased Tumor Uptake In Vivo"
How to order
For details on ordering please visit How to order. Click add to cart above to buy online.
Quotations
Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities or licensing opportunities.
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:
- Immobilized GlycINATOR (EndoS2) hydrolyzes the N-glycans on the Fc domain of the IgG, leaving the innermost GlcNac residue intact.
- Specific attachment of a GalNAz residue to the core GlcNAc by an engineered galactose transferase (GalT(Y298L)).
- 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.
Reliable Performance
Robust method that can be used to modify IgGs of many species and subclasses.
Precise and complete conjugation
Site specific conjugation on the Fc domain preserving the antigen binding activity.
Documents
Contact
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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.
©2018 Genovis AB.