Analytical Sciences

α-Glucosidase Inhibition Assay in an Enzyme-immobilized Amino-microplate

Toshiro MATSUI, Mayu SHIMADA, Nozomi SAITO, and Kiyoshi MATSUMOTO

Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi, Fukuoka 812-8581, Japan

α-Glucosidase (AGH) from the small intestine of rat was immobilized onto a glutaradehyde (GA) activated NH₂-96 well microplate to establish a convenient and rapid AGH inhibition assay system. After AGH immobilization, remaining GA groups were blocked by β-alanine to induce a negative charge on the surface of the well. The AGH-plate showed an enzyme activity of 444 nU/well under an assayed condition at 37°C for 2 h using 0.3 mM 4-methylumbelliferyl-α-D-glucopyranoside as a fluorogenic substrate. Inhibitory powers of voglibose and acarbose as therapeutic AGH inhibitors were successfully evaluated to have IC₅₀ values of 13 and 114 nM, respectively.

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