Investigating oligonucleotide hybridization at subnanomolar level by surface plasmon resonance biosensor method.
We have optimized surface plasmon resonance (SPR) biosensor technology for a rapid, direct, and low-consumption label-free multianalyte screening of synthetic oligonucleotides (ONs) with modified internucleotide linkages potentially applicable in antisense therapy. Monitoring of the ONs hybridization is based on the formation of complex between the natural oligonucleotide probe immobilized on the sensor surface and the ON in solution in contact with the sensor surface. An immobilization chemistry utilizing the streptavidin-biotin interaction was employed to obtain desired ligand density and high hybridization efficiency. It was demonstrated that the sensor is capable of detecting complementary 23-mer ONs in concentrations as low as 0.1 nM with high specificity and reproducibility.
Vaisocherová
Hana
H
Institute of Radio Engineering and Electronics, Academy of Sciences of the Czech Republic, Chaberská 57, 182 51 Prague, Czech Republic. vaisocherova@ure.cas.cz
Zítová
Alice
A
Lachmanová
Markéta
M
Stepánek
Josef
J
Králíková
Sárka
S
Liboska
Radek
R
Rejman
Dominik
D
Rosenberg
Ivan
I
Homola
Jirí
J
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D020349:Surface Plasmon Resonance / Q000379:methods*
D015345:Oligonucleotide Probes