Lectin column

Sepharose-lectin column chromatography To purify glycoproteins or gly-copeptides that bind the particular lectin used. 

Affinity chromatography Has a wide number of uses and can be applied to the isolation and purification of virtually all biomolecules. Specific applications include nucleic acid purification, protein purification from cell and tissues extracts, and antibody purification from blood serum. There are a number of matrices used for the construct, and some examples of these and their uses are as follows heparin columns to separate cholesterol lipoproteins, lectin columns to separate carbohydrate groups, and phenyl boronate columns to separate glycated haemoglobins. 

The affinity elution steps possible—for example, the peptide epitope when using an antibody, imidazole or EDTA when using a His-tag, or specific sugar residue when using a lectin column to purify glycoproteins—are likely to improve the specificity of the isolation process. 

Figure 7. Fluorescence images of arrayed carbohydrates assayed with FITC-labeled lectins. Columns of 4 spots each of 2, 1, 0.5 and 0.1 mM carbohydrates were incubatedfor 20 min with FITC-ConA (top) or FITC-ECA with 1% TWEEN-20 detergent (bottom) with ESA. (Reproduced with permission from reference 19. Copyright 2005 American Chemical Society.)...

The structural information won from HPLC or lectin columns can and should be conflrmed by independent methods. These include NMR, mass spectrometry, and sequencing of purifled oligosaccharide chains with exoglycosidases. 

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