SmartEnzymes


Site-specific conjugation of IgG with azide-alkyne click chemistry.
GlyCLICK Azide Activation contains all reagents needed to azide-activate 250 μg, 2 mg or 10 mg IgG for site-specific custom conjugation using any alkyne-reactive label of choice.
 

  • Site-specific azide activation of IgG with a degree of labeling (DOL) of 2
  • Versatile and consistent custom conjugation of IgG
  • Fc-specific labeling preserves antigen-binding capacity

Available Products

GlyCLICK Azide Activation 250 𝜇gSite-specific conjugation of 250 𝜇g IgG with azide-alkyne click chemistryL1-AZ1-025965.00Buy / Request a Quote
GlyCLICK Azide Activation 2 mgSite-specific conjugation of 2 mg IgG with azide-alkyne click chemistryL1-AZ1-2001,671.00Buy / Request a Quote
GlyCLICK Azide Activation 10 mgSite-specific conjugation of 10 mg IgG with azide-alkyne click chemistryL1-AZ1-1006,696.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

GlyCLICK Azide Activation Citations

Unit Definition

GlyCLICK Azide Activation contains sufficient material for azide activation of 250 μg, 2 mg or 10 mg IgG. For larger amounts or evaluation of the technology, please contact us.

Content and Storage

Components in GlyCLICK Azide Activation:

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

GlyCLICK Azide Activation 250 μg / 2 mg: Content should be stored at +4-8°C upon arrival.
GlyCLICK Azide Activation 10 mg: Content should be stored at different temperatures upon arrival.

FAQ and Support

Popular FAQ

The GlyCLICK 250 ug kit is optimized for 250 ug of IgG as a starting material. We do not recommend using less than 100 ug of starting material in a minimum of 100µL to achieve proper end-over-end mixing on the GlycINATOR columns. Please contact support@genovis.com for assistance if you want to process smaller amounts then 100µg.

The stability of azide activated material varies depending on the antibody and if stabilizing agents have been added. In our labs we have stored azide activated antibodies in TBS buffer without any additives at +4°C for over two years. The azide activation was still OK, and the click reaction generated antibodies with DOL=2. If azide is added as a preservative during long-term storage, it must be removed prior to click reaction.

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.

 

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