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The demand for sensitive and cost-effective diagnostics Analysis is steadily increasing.
Addressable immunoarrays based on matching oligo-deoxynucleotide (ODN) strands have been described before 1,2. Short ODNs (e.g. 10 mers) could be used for addressing but are not stable enough to resist washing steps which lead to a sensitivity reduction.
The demand for sensitive and cost-effective diagnostic tools is steadily increasing. Short ODNs (e.g. 10mers) could be used for addressable immunoarrays 1,2 but are not stable enough to resist washing steps which lead to a sensitivity reduction. Our aim is to establish a new platform for addressable immuno-arrays using shorter ODNs by increasing their stability via covalent linking. The main strategy consisted in the use of furan-modified ODNs that can be selectively oxidized leading to the formation of a site-specific interstrand cross-link (ICL)
Comparison of the surface plasmon resonance (SPR) response shows successful ICL of short DNA duplex on surface. After ICL formation only the binding of the cross-linked DNA duplex with the recognition antibody is stable after regeneration using Na2CO3 (Fig. 1).
Figure1. Method for detection of DNA ICL formation using an FITC labeled ODN and an anti-FITC antibody as model system a) Hybridized ODNs are efficiently separated after using Na2CO3 for regeneration b) Oxidation of the furan using 1O2 results in the formation of a stable ICL, which makes the short ODN duplex stable after regeneration
Were prepared four unsubstituted Pc - C60 fullerene systems, using two different synthetic strategies, via Prato (1,3 - dipolar cycloaddition) or Bingel reactions. The precursor Pcs 1 and 2 were successfully synthesized by cyclotetramerization reaction between TBDPS - protected and unsubstituted phthalonitriles. The surface studies are currently ongoing.
The development of fast and cheap highthroughput platforms for the detection of environmental contaminants is of particular importance to understand the human-related impact on the environment. The application of DNA-directed immobilization (DDI) of IgG molecules is
currently limited to the clinical diagnostics scenario, possibly because of the high costs of production of such addressable platforms. We here describe the efficient and specific hybridization of an antibody−oligonucleotide conjugate to a short 12-mer capture probe. The specific antibody used is a monoclonal antibody against caffeine, a stimulant and important anthropogenic marker. With this work, we hope to contribute to broadening the application potential of DDI to environmental markers in order to develop cheaper and more stable high-throughput screening platforms for standard Routine analysis of pollutants in a variety of complex matrices.