2 Ab Labeling

Glycans are responsible for the structural variations in biological systems. Glycans are labeled at their reducing end using reductive amination. A label containing a primary amine group in a condensation reaction reacts with the aldehyde group of the glycan. The result of this process is a Schiff base, which is reduced by a reducing agent to yield a secondary amine. The most common labels are 2-aminobenzamine (2-AB), 2-aminobenzoic acid (2-AA) and 2-aminopyridine (PA). The 2-AB label lacks negative charges and is used extensively in chromatographic analysis. Labeling agents are usually present in excess hence most of the analytical techniques have to go through a sample workup to remove excess labeling reagents.

Europa Bioproducts Ltd. has a Gly-XTM N-Glycans Rapid Release and Labeling with 2-AB ExpressTM kit which makes use of a unique in-solution enzymatic protein de-glycolisation. N-Glycans are labeled with 2-AB dye after a mere de-glycosylation process carried out in solution. The method is swift and suitable for automation. A clean-up process is then carried out then the labeled glycan samples are ready for analysis. The HPLC method used involves separation by ion exchange, normal phase or size exclusion chromatography. Labeled glycans are usually undetected by UV detection, but through fluorescence, significant peaks can be observed. Mass spectrometry can also detect the labeled glycans. Different chromatograms come up due to the labeling efficiency and sensitivity and dye properties.

For the de-glycolisation, GX96-100 Gly-X De-Glycolisation Module (96-ct) is used, for the labeling process, GX96-401 Gly-X 2-AB Express Labeling Module (96-ct) is used, and as for the last process, the clean-up process, GX96-402 Gly-X 2-AB Express Clean-up Module (96-ct) is used. Other benefits of the Gly-X 2-AB Express include reductive amination 2-AB Express labeling chemistry without dry down, and optimized cleanup removes excess dye, protein, and other interfering compounds.

 

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