Instrumentation for the Interfacial Analysis for Biosensor Microsystems Containing Genetically Engineered Proteins

摘要

The combined use of an atomic force microscope and ellipsometer allows determination of the structure of immobilized protein layers (thickness, orientation, surface concentration, homogeneity), kinetics of layer formation, transport properties of ligand between bulk and protein layer, and stability of the protein layer as compared to solution stability. We have characterized monolayers formed by covalent attachment of E. coli periplasmic binding proteins to glass and other substrates, and found that the orientation of these proteins can be controlled precisely, and that the monolayers retain their ability to respond to analytes with a ligand-mediated hinge-bending conformational change.