An ON/OFF biosensor based on blockade of ionic current passing through a solid-state nanopore.
Single nanopores have attracted interest for their use as biosensing devices. In general, methods involve measuring ionic current blockades associated with translocation of analytes through the nanopore, but the detection of such short time lasting events requires complex equipment and setup that are critical for convenient routine biosensing. Here we present a novel biosensing concept based on a single nanopore in a silicon nitride membrane and two anchor-linked DNA species that forms trans-pore hybrids, realizing a stable blockade of ionic current through the pore. Molecular recognition events affecting the DNA hybrids cause a pore opening and the consequent establishment of an ionic current. In the present implementation of the device, we constructed a magnetic bead/streptavidin/biotin-DNA1/DNA2-biotin/streptavidin/Quantumdot-cluster complex (where DNA1 is a mismatched reverse complement of DNA2) through a sub-micrometric pore and monitored DNA strand displacement events occurring after addition of an oligonucleotide complementary to DNA2. The electric and mechanical aspects of the novel device, as well as its potential in biosensing are discussed.
Dipartimento di Biologia e Protezione delle Piante, Università di Udine, Udine, Italy.
Manola
M
Moretti
Enzo
E
Di Fabrizio
Stefano
S
Cabrini
Rita
R
Musetti
Francesco
F
De Angelis
Giuseppe
G
Firrao
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004247:DNA / Q000032:analysis; D004586:Electrophoresis; D007473:Ion Channels / Q000037:antagonists & inhibitors* / Q000502:physiology; D008280:Magnetics; D008856:Microscopy, Fluorescence; D049329:Nanostructures
D007473:Ion Channels; D004247:DNA