Labeless AC impedimetric antibody-based sensors with pgml(-1) sensitivities for point-of-care biomedical applications.
This paper describes the development and characterisation of labeless immunosensors for (a) the cardiac drug digoxin and (b) bovine serum albumin (BSA). Commercial screen-printed carbon electrodes were used as the basis for the sensors. Two methods were used to immobilise antibodies at the electrode surface. Aniline was electropolymerised onto these electrodes to form a thin planar film of conductive polyaniline; the polyaniline film was then utilised as a substrate to immobilise biotinylated anti-digoxin using a classical avidin-biotin affinity approach. As an alternative approach, poly(1,2-diaminobenzene) was electrodeposited onto the carbon electrodes and this modified surface was then sonochemically ablated to form an array of micropores. A second electropolymerisation step was then used to co-deposit conductive polyaniline along with antibodies for BSA within these pores to produce a microarray of polyaniline protrusions with diameters of several mum, containing entrapped anti-BSA. The resulting antibody grafted electrodes were interrogated using an AC impedance protocol before and following exposure to digoxin or BSA solutions, along with control samples containing a non-specific IgG antibody. The impedance characteristics of both types of electrode were changed by increasing concentrations of antigen up to a saturation level. Calibration curves were obtained by subtraction of the non-specific response from the specific response, thereby eliminating the effects of non-specific adsorption of antigen. Both the use of microelectrode arrays and affinity binding protocols showed large enhancements in sensitivity over planar electrodes containing entrapped antibodies and gave similar sensitivities to our other published work using affinity-based planar electrodes. Detection limits were in the order of 0.1ngml(-1) for digoxin and 1.5ngml(-1) for BSA.
Barton
Andrew C
AC
Cranfield Health, Cranfield University, Silsoe, Beds MK45 4DT, UK.
Collyer
Stuart D
SD
Davis
Frank
F
Garifallou
Goulielmos-Zois
GZ
Tsekenis
Georgios
G
Tully
Elizabeth
E
O'Kennedy
Richard
R
Gibson
Tim
T
Millner
Paul A
PA
Higson
Séamus P J
SP
D000818:Animals; D000906:Antibodies / Q000097:blood*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002417:Cattle; D004147:Dioxins / Q000097:blood*; D017097:Electric Impedance; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D019095:Point-of-Care Systems; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000032:analysis*; D013194:Staining and Labeling
D000906:Antibodies; D004147:Dioxins; D012709:Serum Albumin