Directed immobilization of nucleic acids at ultramicroelectrodes using a novel electro-deposited polymer.
A two-step method for the directed immobilization of nucleic acids at ultramicroelectrodes with micron-size dimensions is described. The approach is based on the immobilization of streptavidin at the surface of carbon or noble metal electrodes within a novel electro-deposited polymer, formed by electropolymerization of the natural compound scopoletin (7-hydroxy-6-methoxy-coumarin) at potentials between 0.4 and 0.7 V vs. Ag/AgCl. Biotin-tagged nucleic acids or proteins are immobilized on top of the modified electrodes in a second step. The new method has some advantages compared to classical electropolymerization approaches (e.g. polypyrrole, polyphenol), because the growing polymer is highly hydrophilic, resulting in efficient incorporation of streptavidin and a high biotin binding capacity of 6 pmol cm(-2). The polymer film seems to be non-conductive but shows good swelling properties in aqueous solutions. The feasibility of the method for the electro-directed biochemical modification of individual microelectrodes has been demonstrated by sequential immobilization of two different single strand oligonucleotides onto interdigitated ultramicroelectrodes. The resulting miniature DNA probe was used for single base mutation detection with two synthetic targets (fluorescence-labeled 17-mer oligomers) by evaluating the fluorescence patterns after hybridisation with the immobilised DNA probes. The new method is useful for the production of microelectrode based DNA chips and for the electro-directed immobilisation of biomolecules at microelectrode structures with high spatial resolution and yield.
Gajovic-Eichelmann
Nenad
N
Fraunhofer Institute for Biomedical Engineering, A-Scheunert-Allee 114-116, 14558 Bergholz-Rehbrucke, Germany. nenad.gajovic@ibmt.fraunhofer.de
Ehrentreich-Förster
Eva
E
Bier
Frank F
FF
D000327:Adsorption; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis* / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004572:Electrolysis / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D008839:Microelectrodes; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods; D011108:Polymers / Q000737:chemistry; D015203:Reproducibility of Results; D012603:Scopoletin / Q000737:chemistry*; D012680:Sensitivity and Specificity
D020099:Coated Materials, Biocompatible; D011108:Polymers; D004247:DNA; D012603:Scopoletin