Figure 1.

A 96-well plate based colorimetric high-throughput screening method for sialidase substrate specificity studies. Sialoside libraries containing sialosides with different sialic acid forms, different sialyl linkages, and different penultimate monosaccharides were incubated in triplicate with different sialidases and an excess amount of a β-galactosidase or a hexosaminidase at 37 °C for 20–60 min. The reaction was stopped by adding CAPS buffer to adjust pH to 9.6. The formation of the p-nitro phenolate was monitored by measuring the A_{405 nm} using a microplate reader. The absorbance of the solution correlates to the amount of the sialic acid cleaved by the sialidase, and to the hydrolytic activity of the sialidase.