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GlyCLICK®

GlyCLICK is a site-specific conjugation technology for IgG using enzymatic remodeling of the Fc-glycans and click-chemistry.

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

The modular format of the technology offers reliable performance with intact immunoreactivity for sensitive applications. Usage of GlyCLICK is versatile and include in vitro and in vivo imaging as well as immunoassays and cytotoxic drug development. The technology is available in a range of kit formats to facilitate tailored and site-specific labeling of antibodies for a diverse range of applications.

GlyCLICK product box

Alexa Fluor 488

In vitro cell imaging, IHC, FCM
250μg (L1-F01-025) $985.00
2mg (L1-F01-200) $1,545.00

Biotin

Immunoassays, ELISA, WB
250μg (L1-A01-025) $985.00
2mg (L1-A01-200) $1,545.00

DFO

Immuno- and In vivo Imaging
250μg (L1-C01-025) $985.00
2mg (L1-C01-200) $1,545.00

Azide Activation

Custom Conjugation
250μg (L1-AZ1-025) $875.00
2mg (L1-AZ1-200) $1,355.00
10mg (L1-AZ1-100) $6,195.00

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:

GlyCLICK workflow visualization

Figure 1.Schematic presentation of the GlyCLICK conjugation process.

  1. Deglycosylation
    Endoglycosidase GlycINATOR® (EndoS2)1 specifically digests IgG Fc-glycans to the inner most GlcNAc moiety on several IgG subclasses and species. The enzyme removes all glycoforms, including; high-mannose, hybrid, complex, and bisecting type glycans. GlyCINATOR is immobilized on spin columns for rapid digestion at room temperature, resulting in a homogenous sample free from any residual enzyme. Learn more about GlycINATOR.
  2. Azide Activation
    Azide-containing UPD-GalNAz is enzymatically attached to the exposed GlcNAc on the Fc-glycans using the ß-1,4-Galactosyltransferase Y289L* (GalT) to generate azide-activated antibodies. The incorporation of an azide at the Fc-glycan sites renders the antibody reactive with an alkyne-containing (cyclooctyne-functionalized) labels in the subsequent conjugation using click-chemistry.
  3. CLICK Reaction
    Azide activated antibodies can then react with cyclooctyne-functionalized labels2 through strain-promoted copper-free click chemistry (SPAAC)3, utilizing the alkyne-azide reaction to form a stable triazole. The biocompatibility of the GlyCLICK technology permits spontaneous and biorthogonal reactions with cyclooctyne-functionalized labels such as fluorophores, biotins, chelators and drug payloads.

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.

Characteristics

GlyCLICK enables site-specific and quantitative conjugation of IgG from several species and subclasses with a degree of label (DOL=2.0). 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. Further more, robust GlyCLICK protocol results in antibody conjugates that are stable and fully labeled thus minimizing the variation between batches.

 

GlyCLICK Characteristics:

 

Key Characteristics

  • Human IgG1-4, Fc-fusion proteins, IgG from mouse, rabbit, rat, monkey, sheep, goat, cow and horse
  • ~2-3 day protocol
  • Available conjugates: AlexaFluor®488, biotin and DFO. Azide activation kits are available for custom conjugation

Applications

The GlyCLICK technology offers a unique and robust method for site-specific conjugation of antibodies. Primary labeled antibodies have applications in many fields research and biopharma development. The ability to conjugate IgG with a wide variety of functionalized conjugates enables a broad range of applications such as in vitro and in vivo imaging as well as immunoassays and cytotoxic drug development.

Fluorescent image

Fluorescent Imaging

Site-specific GlyCLICK conjugation for tailored conjugates suitable for in vitro cell imaging, IHC and FCM.

GlyCLICK ADC development

ADC Development

GlyCLICK generates homogenous ADCs with increased circulation time and minor drug loss.

Immuno imaging <em/>in vivo

In vivo Immuno Imaging

Preserved affinity and increased tumor uptake in vivo for reproducible imaging results using GlyCLICK.

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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.

GlyCLICK characters

GlyCLICK contains

  • Immobilized GlycINATOR
  • UDP-GalNAz
  • GalT(Y298L)
  • sDIBO label of choice
  • All buffers needed
  • Concentration and desalting columns

Unit definition

GlyCLICK contains 250 units for conjugation of 250 μg of antibody. The azide activation kit is also available for conjugation of 2 or 10 mg of antibody. For larger amounts or for evaluation of GlyCLICK for ADC projects, please contact us using the contact form below.

 

Latest references using this Product

Download GlyCLICK Posters

Genvois posters on characteristics and applications for the GlyCLICK technology.

All GlyCLICK References

All GlyCLICK References

Published research articles on GlyCLICK and applications for the technology.

Download GlyCLICK Brochure

Download GlyCLICK Brochure

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

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Legal and Disclaimers

All rights reserved. Genovis products are covered by one or more patents, pending patent applications, trademarks and/or copyrights owned or controlled by Genovis AB.

For more information about commercial rights, please contact Genovis team at info@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

 

SiteClick™ included in GlyCLICK®

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, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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