Silica-immobilized enzyme reactors; application to cholinesterase-inhibition studies.
A rapid and economical method is reported for the preparation of an immobilized enzyme reactor (IMER) using silica-encapsulated equine butyrylcholinesterase (BuChE) as a model system. Peptide-mediated silica formation was used to encapsulate BuChE, directly immobilizing the enzyme within a commercial pre-packed column. The silica/enzyme nanocomposites form and attach simultaneously to the metal affinity column via a histidine-tag on the silica-precipitating peptide. BuChE-IMER columns were integrated to a liquid chromatography system and used as a rapid and reproducible screening method for determining the potency of cholinesterase inhibitors. The IMER preparation method reported herein produces an inert silica-encapsulation matrix with advantages over alternative systems, including ease of preparation, high immobilization efficiency (70-100%) and complete retention of activity during continuous use.
Luckarift
Heather R
HR
Air Force Research Laboratory, Airbase Technologies Division, 139 Barnes Drive, Suite #2, Tyndall Air Force Base, FL 32403, USA. hluckarift@gtcom.net
Johnson
Glenn R
GR
Spain
Jim C
JC
D000818:Animals; D002091:Butyrylcholinesterase / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000032:analysis*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006736:Horses; D008855:Microscopy, Electron, Scanning; D012822:Silicon Dioxide
D002800:Cholinesterase Inhibitors; D004800:Enzymes, Immobilized; D012822:Silicon Dioxide; D002091:Butyrylcholinesterase