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.