Immobilized enzyme reactors in HPLC and its application in inhibitor screening: A review

Abstract

This paper sets out to summarize the literatures based on immobilized enzyme bio-chromatography and its application in inhibitors screening in the last decade. In order to screen enzyme inhibitors from a mass of compounds in preliminary screening, multi-pore materials with good biocompatibility are used for the supports of immobilizing enzymes, and then the immobilized enzyme reactor applied as the immobilized enzyme stationary phase in HPLC. Therefore, a technology platform of high throughput screening is gradually established to screen the enzyme inhibitors as new anti-tumor drugs. Here, we briefly summarize the selective methods of supports, immobilization techniques, co-immobilized enzymes system and the screening model.

Keywords: Immobilized enzyme reactors (IMERs); Molecular bio-chromatography; Screening enzyme inhibitors.

Figure 1

Scheme for the chemical modification of the monolithic silica column using carboxymethyl chitosan. Reproduced from Ref. .

Figure 2

Reaction schemes of covalent immobilization of enzymes with the following activating agents: (a) glutaraldehyde, (b) DSC and (c) DSS.

Figure 3

Reaction schemes of some methods for covalent immobilization of enzymes (a) β-Hydroxylamine formation, (b) Shiff's base formation and (c) amine formation via nucleophilic substitution of tresyl group.