Functionalization of PVC membrane with ss oligonucleotides for a potentiometric biosensor.
A novel application of a single stranded (ss) oligonucleotide as an active component of polymeric membrane in an ion-selective electrode (ISE) is described. The original oligonucleotides, oligo(dA)(15), modified by cholesterol, triphenylmethyl and hexadecyl derivatives, were immobilized into poly(vinyl chloride) (PVC) membrane using extraction protocol. In parallel, the adsorption protocol was used to immobilize unmodified oligo(dA)(15) on the PVC membrane based on tridodecylmethyammonium chloride (TDDMA(+)Cl(-)). Immobilization of ss oligonucleotide probe through spacer was more effective for the potentiometric detection of the hybridization between complementary oligonucleotides. It was found that cholesterol-oligo(dA)(15) modified membranes were sensitive toward complementary oligo(dT)(15) in the concentration range 2-80 nM at pH 7. An explanation for the detection mechanism is proposed.
Shishkanova
T V
TV
Department of Analytical Chemistry, Institute of Chemical Technology, Technická 5, 166 28 Prague 6, Czech Republic. tatiana.shishkanova@vscht.cz <tatiana.shishkanova@vscht.cz>
Volf
R
R
Krondak
M
M
Král
V
V
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D017403:In Situ Hybridization / Q000295:instrumentation* / Q000379:methods; D009841:Oligonucleotides / Q000032:analysis* / Q000737:chemistry* / Q000235:genetics; D011143:Polyvinyl Chloride / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D020099:Coated Materials, Biocompatible; D015342:DNA Probes; D009841:Oligonucleotides; D011143:Polyvinyl Chloride