The Ludger EC50 plate has been designed to purify glycans from non-carbohydrate material including salts, proteins and detergents by electronic interaction of the glycans with the surface of the cartridge.
• Removes salts, detergents, and proteins to enhance HPLC and mass spec analysis
• Easy to use
• Improves the signal intensity after labeling
96-well Ludger EC50 plates can be used:
• After enzymatic or chemical release of glycans from glycoproteins
• After exoglycosidase (enzymatic) digestion of glycans to release individual monosaccharides to confirm glycan identity and structure
• Before and after glycan labelling using fluorescent tags such as 2-aminobenzamide acid (2-AB) or 2-aminobenzoic acid (2-AA)
The 96-well Ludger EC50 plates contain an electron interaction resin (EIR). This unique material contains very flat regions of pi-orbital electrons that interact with hydrophilic organic compounds including glycans.
The resin can capture a wide range of glycans from complex mixtures while salts, proteins, and certain detergents pass through. The glycans are then eluted from the cartridge using a simple solvent system.
2-AB Labeling Efficiency: Use of EC50 Cartridges vs No Clean-up
UHPLC HILIC profile of 2-AB labeled human IgG standards in PBS – 11 replicates;
Black: 2-AB labeling after EC-50 clean-up. Blue: 2-AB labeling with no clean-up performed.
Average absolute area of the human IgG peaks (11 replicates) with EC50 clean-up (black)
and with no clean-up (blue) followed by 2-AB labeling/T1 clean-up
Ludger EC50 Cartridge Reproducibility
UHPLC HILIC profile of 2-AB labeled human IgG – 11 replicates; post EC-50 clean-up of CLIBN-IGG-5U in PBS
EC50 and EB10 Equivalence
Average relative % of human IgG standard peaks post EB10 (8 replicates)
and post EC50 (11 replicates) clean-up comparison
Average relative area % of human IgG standard peaks post EB10 (8 replicates) and post EC50 (11 replicates)
followed by 2-AB labeling and post-labeling cleanup (T1 cartridges)
Binding Capacity Each EC50 cartridge can typically bind up to 50 μg of O- or N-linked glycans.
Number of Samples Ludger EC50 plates can be used with up to 96 samples.
Suitable Samples A wide range of glycans can be purified. These include N-linked and O-linked type oligosaccharides, tri-saccharides and larger structures.
The cartridges are not suitable either for monosaccharides or disaccharides which are generally bound too weakly for efficient purification or for large linear poly-sialylated glycans which can be bound very tightly to the resin.
Glycan samples must be applied to the cartridges in solutions that are substantially aqueous.
Structural Integrity No detectable (< 2 mole per cent) loss of sialic acid, fucose, sulfate, or phosphate.
Binding efficiency >95% for most glycans
Binding Selectivity Essentially stoichiometric binding and elution for most complex glycan mixtures.
Storage Store at room temperature in the dark. Protect from sources of heat, light, and moisture. The cartridges are stable for at least two years as supplied.
Shipping The product can be shipped at ambient temperature.
Handling Ensure that any glass, plastic ware or solvents used are free of glycosidases and environmental carbohydrates. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrate.
Safety Please read the Safety Data Sheet (SDS) for all chemicals used.
All processes involving hazardous reagents should be performed using appropriate personal safety protection – eyeglasses, chemically resistant gloves (e.g. nitrile), etc. – and where appropriate in a laboratory fume hood.