NEW! Azide Activation of Human IgG using TransGLYCIT Azide Activation

We’re happy and proud to announce the launch of TransGLYCIT Azide Activation!
 

The new TransGLYCIT Azide Activation product enables efficient and site-specific conjugation at the Fc N-linked glycosylation site of human IgG. The azide activation is performed within only a few hours, and the resulting antibody can be labeled with any click reagent of choice to a degree of labeling (DOL) of four labels per antibody. A site-specific labeling with a high DOL is desirable for fluorescent-based assays, since it allows for a high sensitivity. In antibody-drug conjugates, the drug-antibody ratio impacts the potency. Also, the TransGLYCIT technology ensures quantitative conjugation and intact immunoreactivity, features that are often limited when using random conjugation technologies.

TransGLYCIT Azide Activation was here used for site-specific azide activation of trastuzumab using a modified N-glycan oxazoline carrying two azide functionalities. The reaction generated an IgG with four sites for conjugation, two sites on each Fc/2, as shown in the figure below.

 
Azide activation of trastuzumab by the TransGLYCIT workflow. Deconvoluted mass spectra of the Fc/2 fragment of native trastuzumab (top), after deglycosylation by GlycINATOR (middle) and after transglycosylation by TransINATOR with the oxazoline glycoform azide as the substrate (bottom). The mAb was digested with FabRICATOR and the subunits were analyzed by reversed-phase LC-MS on a Waters™ BioAccord™ system equipped with a Waters™ BioResolve™ RP mAb column (2.1×50mm).

TransGLYCIT Azide Activation – Azide activation of 100 µg human IgG1 or IgG4

TransGLYCIT Azide Activation hIgG2 – Azide activation of 100 µg human IgG2