Screen printing of nucleic acid detecting carbon electrodes.
A large fraction of the presently mass-manufactured (> 10(8) units/year) electrochemical biosensors, used mostly by diabetic people to monitor their blood glucose levels, have screen-printed carbon working electrodes. An earlier study (Campbell, C. N., et al. Anal. Chem. 2002, 74, 158-162) showed that nucleic acids can be assayed at 1 nM concentrations by a sandwich-type amperometric method. The assay was performed with vitreous carbon working electrodes on which an electron-conducting polycationic redox polymer and avidin were coelectrodeposited. Because the rate of the electrodeposition increases with the surface density of the polycationic redox polymer, its practicality depends on pretreatment of the surface, which adds anionic functions. (Gao, Z., et al. Angew. Chem. Int. Ed. 2002, 41, 810-813). Here it is shown that the required conducting redox polymer films can be electrodeposited on potentially mass manufacturable electrodes made by screen-printing hydrophilic carbon inks on polyester sheets. The modified electrodes are made in two steps. First a polycationic electron-conducting redox polymer is cross-linked and electrodeposited by applying a negative potential. Next, an amine-terminated 20-base single-stranded oligonucleotide is electrodeposited by ligand-exchange. Both steps involve exchange of a labile inner sphere chloride ligand of the polymer-bound osmium-complex: Cross-linking and electrodeposition of the redox polymer result when inner-sphere chloride anions of the osmium complexes are exchanged by imidazole functions of neighboring chains. Incorporation of the oligonucleotide in the redox polymer results in the formation of a coordinative bond between the terminal amine (attached through a spacer to the oligonucleotide) and the osmium complex. In testing for the presence of a 38-base oligonucleotide, the analyte, in a 15- or 25-microL droplet of hybridization solution, is hybridized with and captured by the 20-base electrode-bound sequence; then it is hybridized with an 18-base horseradish peroxidase labeled sequence. When the HRP label electrically contacts the redox polymer, the film becomes an electrocatalyst for the reduction of H2O2 to water at 0.10 V (Ag/AgCl). Flow of the H2O2-reduction current indicates the presence of the assayed sequence.
Dequaire
Murielle
M
Department of Chemical Engineering and Texas Materials Institute, The University of Texas at Austin, 78712, USA.
Heller
Adam
A
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D002244:Carbon; D004566:Electrodes / Q000592:standards; D009696:Nucleic Acids / Q000032:analysis*; D012680:Sensitivity and Specificity; D013499:Surface Properties
D009696:Nucleic Acids; D002244:Carbon