title: Immobilisation of DNA probes for the development of SPR-based sensing.
abstract: An immobilisation procedure based on the direct coupling of thiol-derivatised oligonucleotide probes to bare gold sensor surfaces has been used for DNA sensing applications. The instrumentation used relies on surface plasmon resonance (SPR) transduction; in particular the commercially available instruments BIACORE X and SPREETA, have been employed in this study. The performances of the SPR-based DNA sensors resulting from direct coupling of thiol-derivatised DNA probes onto gold chips, have been studied in terms of the main analytical parameters, i.e. selectivity, sensitivity, reproducibility, analysis time, etc. A comparison between the thiol-derivatised immobilisation approach and a reference immobilisation method, based on the coupling of biotinylated oligonucleotide probes onto a streptavidin coated dextran sensor surface, using synthetic complementary oligonucleotides has been discussed. Finally, a denaturation method to obtain ssDNA ready for hybridisation analysis has been applied to polymerase chain reaction (PCR) amplified samples, for the detection of genetically modified organisms (GMOs).
authors.affiliation: Chemistry and Chemical Engineering College, Central South University, Changsha 410082, Hunan, PR China.
authors.forename: Ronghui
authors.initials: R
authors.lastname: Wang
authors.forename: Sara
authors.initials: S
authors.lastname: Tombelli
authors.forename: Maria
authors.initials: M
authors.lastname: Minunni
authors.forename: Maria Michela
authors.initials: MM
authors.lastname: Spiriti
authors.forename: Marco
authors.initials: M
authors.lastname: Mascini
mesh_terms: D000327:Adsorption; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry*; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
chemical_list: D020099:Coated Materials, Biocompatible; D015342:DNA Probes; D004247:DNA