Rapid and label-free screening of enzyme inhibitors using segmented flow electrospray ionization mass spectrometry.
Electrospray ionization mass spectrometry (ESI-MS) is an attractive analytical tool for high-throughput screening because of its rapid scan time and ability to detect compounds without need for labels. Impediments to the use of ESI-MS for screening have been the relatively large sample consumed and slow sample introduction rates associated with commonly used flow injection analysis. We have previously shown that by segmenting nanoliter plugs of sample with air, an array of discrete samples can be delivered to a platinum-coated emitter tip for ESI-MS analysis with throughput as high as 0.8 Hz and carry-over between samples less than 0.1%. This method was applied to screening for inhibitors of acetylcholinesterase as a demonstration of the potential of segmented flow ESI-MS for such applications. Each enzyme assay consumed 10 nL of sample. At 1 microL/min infusion rate, 102 samples were analyzed, corresponding to a 0.65 Hz sample analysis rate. Linear quantification of choline was achieved from 200 microM to 10 mM using this method and Z' values were over 0.8 for the assay. Detailed pharmacologic dose-response curves of selected inhibitors were also measured in high-throughput fashion to validate the method.
Pei
Jian
J
Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
Li
Qiang
Q
Kennedy
Robert T
RT
D000109:Acetylcholine / Q000737:chemistry / Q000378:metabolism*; D000110:Acetylcholinesterase / Q000378:metabolism; D002800:Cholinesterase Inhibitors / Q000737:chemistry / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004491:Edrophonium / Q000737:chemistry / Q000378:metabolism; D016014:Linear Models; D008294:Malathion / Q000737:chemistry / Q000378:metabolism; D009388:Neostigmine / Q000737:chemistry / Q000378:metabolism; D010830:Physostigmine / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D002800:Cholinesterase Inhibitors; D009388:Neostigmine; D004491:Edrophonium; D010830:Physostigmine; D000110:Acetylcholinesterase; D000109:Acetylcholine; D008294:Malathion