DSL contains two chains of 40 kDa and 46 kDa joined by disulfide bonds. This lectin is free of a reported 32 kDa contaminant protein. The carbohydrate binding site recognizes (Î²-1,4) linked N-acetylglucosamine oligomers, preferring chitobiose or chitotriose over a single N-acetylglucosamine residue. This lectin binds well in the acidic pH range but its affinity decreases above pH 8.0.
DSL also binds well to N-acetyllactosamine and oligomers containing repeating N-acetyllactosamine sequences. A branched pentasaccharide including two N-acetyllactosamine disaccharides linked to mannose (Î²-1,6) and (Î²-1,2) was reported to be the most potent inhibitor of agglutination.
Agarose bound* Datura stramonium lectin is prepared using our affinity-purified lectins. Heat stable, cross-linked 4% agarose beads with a molecular weight exclusion limit of about 2x107 daltons are used as the solid-phase matrix to which the lectins are covalently coupled. The attachment of the lectins to the beads is carefully controlled to preserve lectin activity and minimize conformational changes of the bound lectins that might result in nonspecific ionic or hydrophobic interactions. The technique we have developed to couple lectins to agarose beads inserts a hydrophilic spacer arm between the lectin and the matrix.
This coupling method provides several advantages over the traditional cyanogen bromide procedure:
Maximum carbohydrate binding activity of the coupled lectins is retained
Linkage is stable over a range of pH values
Conjugated proteins are not leached off the beads by Tris or other routinely used buffers
No residual charges are present after conjugation. This minimizes non-specific binding to the matrix.
Our agarose bound lectins are supplied at a constant concentration of lectin per ml of settled beads. The concentration for each lectin is selected to achieve the highest glycoconjugate binding capacity per mg of lectin present in the beads. Each lot is tested for its binding capacity using glycoproteins known to bind the lectin. This provides a guideline for the user and assures the quality of our agarose bound lectins.