Monosaccharide Kit

LudgerTag Quantitative Monosaccharide Analysis Kit
Part number LT-MONO-96

This kit contains reagents for the release and fluorescent labeling of monosaccharides found on glycoprotein biopharmaceuticals and standards. One kit contains reagents to label up to 96 separate analytical samples per kit, in two sets of reagents (48 samples per set). Typically, 50 micrograms of glycoprotein is analyzed per sample.

Released monosaccharides have a free reducing terminus allowing fluorescent tagging with 2-AA (2-aminobenzoic acid).

There are two hydrolysis acids provided. 2 molar trifluoroacetic acid (2M TFA) and 6 molar hydrochloric acid (6M HCl). 2M TFA is good for releasing neutral monosaccharides but is less effective with the N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc) monosaccharides for which we recommend using 6M HCl. We recommend using these acids on separate replicates of your samples.

LudgerTag Release Reagents
   Trifluoroacetic acid - 2M TFA
   Hydrochloric acid - 6M HCl

LudgerTag Monosaccharide Standards
   MonoMix monosaccharide standards - 10 nmols each of glucosamine, galactosamine, galactose, mannose and fucose
   Xylose standard - 100 nmols - used as an internal reference standard for accurate quantification

LudgerTag Labeling Reagents
   2-aminobenzoic acid (2-AA dye)
   Dimethyl sulfoxide (DMSO)
   Sodium acetate
   sodium acetate/boric acid/methanol
   Sodium cyanoborohydride

HPLC -Quantitative Analysis of Labelled Monosaccharides

Column: After release and labelling of monosaccharides, the samples are run on a reverse phase HPLC column. This kit has been optimized for use with the LudgerSep R1 Column. For analysis of the 2-AA labeled monosaccharides, the LudgerSep R1 gives very good separation of the seven main monosaccharides found in most N-link and O-link glycans.

Solvents: The glycan analysis gradients in this guide are based on the following solvents:

Solvent A : purified water based solvent of 0.2 % butylamine (2 mL per litre), 0.5 % phosphoric acid 5 mL per litre), 1 % tetrahydrofuran (10 mL per litre) (henceforth called BPT solvent).

Solvent B : 50 % acetonitrile : 50 % solvent A

We have also tested acetonitrile free solvent systems. Please enquire for further details.