An ultrathin platinum film sensor to measure biomolecular binding.
A sensitive conductimetric immunosensor has been demonstrated based on an ultrathin platinum film on an oxidized silicon base. The film is about 25 A thick and is seen to consist of a discontinuous layer with channels 20-30 A wide. Monoclonal antibodies were bound to the sensor surface using conventional biosensor chemistry. Impedance at fixed frequencies across the film was used to track modification and binding at the surface. Impedance increased 55% at 20 Hz during the activation of the surface with anti-alkaline phosphatase (anti-AP). Binding of alkaline phosphatase (AP) to the prepared surface results in a further increase of 12%. p-Nitrophenyl phosphate hydrolysis confirmed binding and activity of the AP. About 40 amol AP were bound on the 0.5 cm(2) electrode. Non-specific binding of horseradish peroxidase caused an impedance change <6%. Control experiments showed small impedance changes and trace enzyme activity. Since the mechanism of electrical conduction of the thin film was not established, modeling of thin-film response was used to distinguish between redox processes, capacitance and tunneling mechanisms. The data fit well with the diffusion distributed elements (DE) model as well as a transmission line distribution element (DX) model. The first model, DE, is distributed elements for diffusion. The second DX model represents a transmission line. The sensors behave in a distributed network or like a transmission line.
Pak
S C
SC
Bioengineering Department, Microfabrication Applications Laboratory, University of Illinois at Chicago, Chicago, IL 60607, USA.
Penrose
W
W
Hesketh
P J
PJ
D000469:Alkaline Phosphatase / Q000276:immunology / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000032:analysis*; D001665:Binding Sites / Q000502:physiology; D001685:Biological Factors / Q000032:analysis*; D015374:Biosensing Techniques; D003217:Conductometry; D017097:Electric Impedance; D006735:Horseradish Peroxidase / Q000378:metabolism; D008958:Models, Molecular; D010984:Platinum; D012680:Sensitivity and Specificity; D013499:Surface Properties
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D001685:Biological Factors; D010984:Platinum; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase