Ultrafast immunoassays by coupling dielectrophoretic biomarker enrichment in nanoslit channel with electrochemical detection on graphene.
Heterogeneous immunoassays usually require long incubation times to promote specific target binding and several wash steps to eliminate non-specific binding. Hence, signal saturation is rarely achieved at detection limit levels of analyte, leading to significant errors in analyte quantification due to extreme sensitivity of the signals to incubation time and methodology. The poor binding kinetics of immunoassays at detection limit levels can be alleviated through creating an enriched analyte plug in the vicinity of immobilized capture probes to enable signal saturation at higher levels and at earlier times, due to higher analyte association and its faster replenishment at the binding surface. Herein, we achieve this by coupling frequency-selective dielectrophoretic molecular dam enrichment of the target biomarker in physiological media to capture probes immobilized on graphene-modified surfaces in a nanoslit to enable ultrafast immunoassays with near-instantaneous (<2 minutes) signal saturation at dilute biomarker levels (picomolar) within ultra-low sample volumes (picoliters). This methodology is applied to the detection of Prostate Specific Antigen (PSA) diluted in serum samples, followed by validation against a standard two-step immunoassay using three de-identified patient samples. Based on the ability of dielectrophoretic molecular dam analyte enrichment methods to enable the detection of PSA at 1-5 pg mL(-1) levels within a minute, and the relative insensitivity of the signals to incubation time after the first two minutes, we envision its application for improving the sensitivity of immunoassays and their accuracy at detection limit levels.
Department of Electrical & Computer Engineering, University of Virginia, Charlottesville, Virginia-22904, USA. nswami@virginia.edu.
Bankim J
BJ
Sanghavi
Walter
W
Varhue
Ali
A
Rohani
Kuo-Tang
KT
Liao
Lindsay A L
LA
Bazydlo
Chia-Fu
CF
Chou
Nathan S
NS
Swami
D055910:Antibodies, Immobilized / Q000737:chemistry; D055664:Electrochemical Techniques / Q000191:economics / Q000295:instrumentation*; D046209:Electrophoresis, Microchip / Q000191:economics / Q000295:instrumentation; D004867:Equipment Design; D005260:Female; D006108:Graphite / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000191:economics / Q000295:instrumentation*; D057230:Limit of Detection; D049329:Nanostructures / Q000737:chemistry*; D017430:Prostate-Specific Antigen / Q000097:blood*
D055910:Antibodies, Immobilized; D006108:Graphite; D017430:Prostate-Specific Antigen