Biotin
Site-specific conjugation of IgG with biotin.
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.
GlyCLICK Biotin is available for site-specific labeling of 250 μg or 2 mg IgG with biotin, and is suitable for immunoassays, ELISA and Western Blot.
- Site-specific biotin-conjugation to IgG with a degree of labeling (DOL) of 2
- Biotinylation while maintaining immunoreactivity
- Supports enhanced signaling of detected target antigens
Available Products
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Unit Definition
GlyCLICK Biotin contains sufficient material for labeling of 250 μg or 2 mg IgG. For larger amounts or evaluation of the technology, please contact us.
Content and Storage
Components in GlyCLICK Biotin:
- GlycINATOR Immobilized
- UDP-GalNAz
- GalT(Y298L)
- sDIBO (Biotin)
- All buffers needed
- Concentration and desalting columns
Content should be stored at different temperatures upon arrival.
Instructions
Safety Data Sheet
Certificate of Analysis
FAQ and Support
Popular FAQ
Problems can arise from freezing of the conjugates. We cannot guarantee the quality after freezing and therefore recommend storage at +4°C.
The final step of GlyCLICK labeling kit is based on a copper free click reaction (SPAAC) and it is a strain promoted reaction between an azide and an alkyne-carrying label. As the name indicates the reaction occurs spontaneously due to the strained bond angles. The use of this click reaction that forms stable conjugates, allows you to freely choose label or payload to combine with your IgG as long as it is alkyne-functionalized. Several alkyne-carrying labels are commercially available with cyclooctyne structures such as DIBO, DBCO or BCN.
All the steps (deglycosylation, azide activation and click reaction) in GlyCLICK are highly efficient. Normally, all molecules are conjugated when using GlyCLICK, resulting in DOL=2. On the application page we have shown by MS-analysis that all molecules have been conjugated and there is no remaining un-conjugated material.
Characterization using LC-MS will give a complete and clear picture of mass shift associated with successful conjugation if the antibody is analyzed reduced or fragmented. LC or LC-MS analysis of HC from a reduced sample or scFc fragments generated by FabRICATOR digestion provides distinct elution peaks for conjugated scFc or mass shifts corresponding to the degree of labelling (DOL) following conjugation. On the application page you can see an example of such analysis. Analysis of fluorescently labeled antibodies can also be performed by absorbance measurements and calculate the DOL using the Extinction coefficient for the antibody and the label and the correction factor for the label.
Support