title: Electrochemical assay of α-glucosidase activity and the inhibitor screening in cell medium.
abstract: An electrochemical method is established in this work for the assay of α-glucosidase activity and the inhibitor screening through one-step displacement reaction, which can be directly used in cell medium. The displacement reaction can be achieved via strong binding of 4-aminophenyl-α-D-glucopyranoside (pAPG)/magnetic nanoparticles (MNPs) to pyrene boric acid (PBA) immobilized on the surface of graphite electrode (GE), compared to that of dopamine (DA)/sliver nanoparticles (AgNPs). Since α-glucosidase can specifically catalyze MNPs/pAPG into MNPs/pAP which has no binding capacity with PBA, the activity of both isolated and membrane bound enzyme can be well evaluated by using this proposed method. Meanwhile, signal amplification can be accomplished via the immobilization of DA at the outer layer of AgNPs, and the accuracy can be strengthened through magnetic separation. Moreover, this method can also be utilized for inhibitor screening not only in the medium containing the enzyme but also in cell medium. With good precision and accuracy, it may be extended to other proteases and their inhibitors as well.
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China.
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China; Shanghai Key Laboratory of Bio-Energy Crops, Shanghai University, Shanghai 200444, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China.
authors.forename: Xiaonan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China.
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Laboratory of Biosensing Technology, School of Life Sciences, Shanghai University, Shanghai 200444, PR China; State Key Laboratory of Pharmaceutical Biotechnology, Department of Biochemistry, Nanjing University, Nanjing 210093, PR China. Electronic address: genxili@nju.edu.cn.
authors.forename: Genxi
authors.initials: G
authors.lastname: Li
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation*; D002460:Cell Line; D003217:Conductometry / Q000295:instrumentation*; D003470:Culture Media / Q000378:metabolism; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000378:metabolism*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D065089:Glycoside Hydrolase Inhibitors / Q000032:analysis*; D058185:Magnetite Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013552:Swine; D000520:alpha-Glucosidases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003470:Culture Media; D065089:Glycoside Hydrolase Inhibitors; D058185:Magnetite Nanoparticles; D000520:alpha-Glucosidases
keywords: Cell; Electrochemical method; Inhibitor; Magnetic separation; One-step displacement reaction; Pyrene boric acid; α-Glucosidase