Immunosensing platforms using spontaneously adsorbed antibody fragments on gold.
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.
O'Brien
J C
JC
Microanalytical Instrumentation Center, Ames Laboratory-USDOE, Iowa, USA.
Jones
V W
VW
Porter
M D
MD
Mosher
C L
CL
Henderson
E
E
D000327:Adsorption; D000818:Animals; D015374:Biosensing Techniques; D000939:Epitopes; D006046:Gold; D007140:Immunoglobulin Fab Fragments; D007150:Immunohistochemistry; D011817:Rabbits; D013438:Sulfhydryl Compounds; D013499:Surface Properties
D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D013438:Sulfhydryl Compounds; D006046:Gold