title: Preparation of a dual-enzyme co-immobilized capillary microreactor and simultaneous screening of multiple enzyme inhibitors by capillary electrophoresis.
abstract: A CE method based on a dual-enzyme co-immobilized capillary microreactor was developed for the simultaneous screening of multiple enzyme inhibitors. The capillary microreactor was prepared by co-immobilizing adenosine deaminase and xanthine oxidase on the inner wall at the inlet end of the separation capillary. The enzymes were first immobilized on gold nanoparticles, and the functionalized gold nanoparticles were then assembled on the inner wall at the inlet end of the separation capillary treated with polyethyleneimine. With the developed CE method, the substrates and products were baseline separated within 3 min. The activity of the immobilized enzyme can be directly detected by measuring the peak height of the products. A statistical parameter Z' factor was recommended for evaluation of the accuracy of a drug screening system. In the present study, it was calculated to be larger than 0.5, implying a good accuracy. Finally, screening a small compound library containing two known enzyme inhibitors and 20 natural extracts by the proposed method was demonstrated. The known inhibitors were identified, and some natural extracts were found to be positive for two-enzyme inhibition by the present method.
authors.affiliation: Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources, Ministry of Education, College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin, PR China.
authors.forename: Pingtan
authors.initials: P
authors.lastname: Lin
authors.forename: Shulin
authors.initials: S
authors.lastname: Zhao
authors.forename: Xin
authors.initials: X
authors.lastname: Lu
authors.forename: Fanggui
authors.initials: F
authors.lastname: Ye
authors.forename: Hengshan
authors.initials: H
authors.lastname: Wang
mesh_terms: D000243:Adenosine Deaminase / Q000737:chemistry / Q000378:metabolism*; D015374:Biosensing Techniques; D019075:Electrophoresis, Capillary / Q000295:instrumentation; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000187:drug effects; D006046:Gold / Q000737:chemistry / Q000378:metabolism; D053768:Metal Nanoparticles / Q000737:chemistry; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013499:Surface Properties; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D004800:Enzymes, Immobilized; D006046:Gold; D011094:Polyethyleneimine; D014969:Xanthine Oxidase; D000243:Adenosine Deaminase
keywords: Adenosine deaminase; CE; Capillary microreactor; Inhibitor screening; Xanthine oxidase