Immunosensing Platforms Using Spontaneously Adsorbed Antibody Fragments on Gold

Abstract

This paper describes the construction and characterization of miniaturized antigenic immunosurfaces composed of spontaneously adsorbed Fab'-SH fragments on gold. Rabbit Fab'-SH fragments contain a free sulfhydryl group that forms a thiolate bond with a gold substrate as detailed by X-ray photoelectron spectroscopy. This approach creates surfaces of higher epitope density, a factor critical to the early detection of disease, than surfaces composed of adsorbed whole molecule IgG on gold. The viability and specificity of antigenic Fab'-SH immunosurfaces is demonstrated using atomic force microscopy and confocal fluorescence microscopy, and possible explanations for the larger epitope density are discussed.