Site-specific Antibody Conjugation
GlyCLICK is a site-specific conjugation technology for glycan-mediated labeling of native IgG. The technology is based on a unique combination of enzymes and click chemistry that enables Fc glycan remodeling and incorporates two labels per antibody for a DOL/DAR 2.0.
The reliable performance facilitates quantitative labeling and maintains intact immunoreactivity. The GlyCLICK technology can be used with the label or payload of choice for consistent labeling of antibodies intended for different applications.
Advantages:
- Conjugation of native IgG without engineering
- Consistent degree of labeling (DOL) of 2.0
- Preserves affinity and reduces background sign
Deliverables:
- Conjugation of native IgG without engineering
- Consistent degree of labeling (DOL) of 2.0
- Preserves affinity and reduces background sign
Click on graphic to enlarge
Figure 5. HER2(+) cells or tissue. Left: PFA-fixed cells incubated with DAPI (0.2 uM), GlyCLICK-AlexaFluor647 (1 ug/ml). Middle: PFA-fixed cells incubated with DAPI (0.2 uM), GlyCLICK-AlexaFluor647 (5 ug/ml), phallodinAF488. Right: paraffin embedded, BSA blocked tissue incubated with DAPI (0.2 uM), GlyCLICK-AlexaFluor647 (10 ug/ml).
Applications
The ability to conjugate various species and subclasses of IgG using a functionalized label of choice enables a wide range of applications including immuno imaging, bioassays and ADC development.
Fluorescent Conjugates
Site-specific antibody conjugates using various fluorescent dyes for a wide range of imaging applications.
Custom Conjugates
Tailored conjugates using a custom label or payload for homogenous and robust antibody conjugates.
Radiolabeled Conjugates
Quantitative conjugation for high quality chelation and radio-labeling of antibodies for in vivo imaging.
How GlyCLICK works
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Figure 1. Schematic presentation of the GlyCLICK conjugation process.
1. Deglycosylation
The Endoglycosidase GlycINATOR® (EndoS2)1 hydrolyzes the Fc glycans to the inner most GlcNAc moiety on several subclasses and species of IgG. The enzyme can trim all Fc glycoforms, including; high-mannose, hybrid, complex, and bisecting type glycans.
2. 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.
3. CLICK Reaction
The azide activated antibody is conjugated with a functionalized label2 of choice in a biorthogonal reaction through strain-promoted copper-free click chemistry (SPAAC)3 to form a stable triazole.
GlyCLICK Antibody Labeling
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Label Your Own Antibody using GlyCLICK
The GlyCLICK technology is available in a range of easy-to-use kit formats to facilitate tailored conjugation by site-specific labeling of antibodies using customized labels.
Enzymes for Biopharmaceuticals
Genovis provides unique enzymes and technologies for analytical characterization and conjugation of biopharmaceuticals. The SmartEnzymes™ portfolio consists of FabRICATOR® (IdeS), FabRICATOR®Z (IdeZ), FabALACTICA® (IgdE), FabULOUS® (SpeB), GingisKHAN® (KGP), GingisREX® (Rgp), IgGZERO® (EndoS), GlycINATOR® (EndoS2), OpeRATOR®, GlycOCATCH™, OglyZOR®, SialEXO® and the antibody conjugation technology GlyCLICK®.