Glycan derivatization

The general form for oligosaccharide profiling is release of the glycans, derivatization (e. g. fluorescent labeling), then analysis by HPLC, CE, or MS. 

D. J. Harvey, Collision-induced fragmentation of negative-ions from N-hnked glycans derivatized with 2-aminobenzoic acid, 7. Mass Spectrom. 40, 642-655 (2005). 

Okafo, G., J. Langridge, S. North et al. 1997. High-performance liquid chromatographic analysis of complex A-linked glycans derivatized with 2-aminoacridone. Anal. Chem. 69 4985-4993. 

Add to the glycan solution the molecule to be labeled containing an available amine, hydrazine, or hydrazide group. For small molecule derivatization, the final concentration of the nucleophile in the glycan solution should be about 0.3 M to result in maximal efficiency of labeling. For protein modification, an aqueous reaction buffer should be used, and the protein should be as concentrated as possible. 

Rice, K.G. (2000) Derivatization strategies for preparing N-glycan probes. Anal. Biochem., 283, 10-16. 

L. A. Gennaro, D. J. Harvey, and P. Vouros, Reversed-phase ion-pairing liquid chromatog-raphy/ion trap mass spectrometry for the analysis of negatively charged, derivatized glycans, Rapid Common. Mass Spectrom., 17 (2003) 1528-1534. 

Zhuang et al. [152] derivatized glycan samples with 8-aminopyrene-l,3,6-trisulfonic acid to get a charge for electrophoresis and a fluorescent label for detection. They analyzed glycan derivatives in the blood of cancer patients... 

An elegant alternative approach for noncovalent interaction relies on fluorous-lluorous interactions. A glycan array of monosaccharides and disaccharides bearing anomeric fluorous tags was noncovalently immobilized on fluorous-derivatized glass slides (19, 20). The attachment method is compatible with a wide range of functional groups and has been successfully used to probe carbohydrate-protein interactions. 

Xia B, Kawar ZS, Ju T, Alvarez RA, Sachdev GP, Cummings RD. Versatile fluorescent derivatization of glycans for glycomic analysis. Nat. Methods 2005 2 845-850. 

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