Combinatorial Approach to Study Enzyme/Surface Interactions

Published

March 1, 2005

Author(s)

K Loos, S L. Kennedy, Naomi Eidelman, Y Tai, M Zharnikov, Eric J. Amis, A Ulman, R A. Gross

Abstract

A fast combinatorial approach to access information on the immobilization behavior and kinetics of enzymes on a variation of surfaces is presented. As a test system, Candida Antarctica Lipase B (CAL-B) was immobilized on a self-assembled monolayer (SAM) bearing a gradient of surface energy. The respective immobilization behavior was monitored by FTIR microspectroscopy. In addition, the activity of the immobilized enzyme was monitored over the entire film in real time with a specially developed fluorescence activity assay embedded into a siloxane gel. It was found that the highest amount of active protein was immobilized on the hydrophilic end of the gradient surface. This effect is presumably associated with a higher surface roughness of this area resulting in hydrophobic microenviroments in which the enzyme gets immobilized.

Citation

Langmuir

Volume

21

Pub Type

Journals

Keywords

Candida Antartica Lipase B, enzyme catalyst, fluorescence activity assay, FTIR-Microspectroscopy, gradient surfaces, immobilization

Citation

Loos, K. , Kennedy, S. , Eidelman, N. , Tai, Y. , Zharnikov, M. , Amis, E. , Ulman, A. and Gross, R. (2005), Combinatorial Approach to Study Enzyme/Surface Interactions, Langmuir (Accessed April 19, 2024)