| AVS 54th International Symposium | |
| Plasma Science and Technology | Thursday Sessions |
| Session PS2+BI-ThA |
| Session: | Plasmas in Bioscience |
| Presenter: | P. Rivolo, Politecnico di Torino, Italy |
| Authors: | P. Rivolo, Politecnico di Torino, Italy S. Lo Bartolo, LaTEMAR, Ctr of Excellence funded by MIUR; Biodiversity SpA, Italy D. Perrone, Lab. Materiali e Microsistemi, Italy; Politecnico di Torino, Italy S. Fiorilli, LaTEMAR, Ctr of Excellence funded by MIUR; Politecnico di Torino, Italy I. Vallini, LaTEMAR, Ctr of Excellence funded by MIUR; Biodiversity SpA, Italy C. Ricciardi, LaTEMAR, Ctr of Excellence funded by MIUR; Politecnico di Torino, Italy M. Quaglio, Lab. Materiali e Microsistemi, Italy; Politecnico di Torino, Italy G. Mantero, Biodiversity SpA, Italy C.F. Pirri, LaTEMAR, Ctr of Excellence funded by MIUR; Politecnico di Torino, Italy |
| Correspondent: | Click to Email |
The low cost of production, the easy handling and the large variety of polymeric materials favour them as attractive candidates to replace classic glass slides in micro-array biomolecular diagnostics. However, the lack of reactive functional groups, at polymeric surfaces, makes difficult their use as substrates for immobilization of molecules such as DNA fragments, in either cDNA or oligodeoxyribonucleotide (ODN) format, for a variety of applications to DNA micro-array technology including microscale sequencing, mRNA expression monitoring and single nucleotide polymorphism analysis. In this contribution, modification of surface chemical properties of cyclo olefin copolymer (COC), polystyrene (PS), polyethylene (PE) and polycarbonate (PC) is reported. The surface of polymeric substrates, properly molded by hot embossing, was modified introducing monotype functional groups1 and using them for subsequent covalent grafting of linker molecules, active for amino-oligonucleotide probes immobilization.2 A first step consisting of non-equilibrium low-pressure air and water RF plasma was used to activate the polymer surface, forming oxidized species such as -C-O-, -C=O, -C-OH, -CHO, -COOH. Successively, a liquid-phase reduction by a NaBH4 solution was carried out to increase the yield of –OH groups in order to enhance the amount of covalently grafted 3-aminoproylsilane (3-APTES), a reaction carried out by vapour-phase process.3 The last step was performed by a liquid phase reaction between glutaraldehyde and amino-groups of grafted 3-APTES. After this, polymer surface shows -CHO species suitable for the reaction with the amino-modified probes. Characterization of the functionalised polymeric surfaces was performed by contact angle measurements and reflection-absorption infrared spectroscopy (RAIRS)4. Modification efficiency of different polymers substrates was evaluated by well-modified Arrayed Primer EXtension (APEX) protocol with colorimetric and fluorimetric detection methods.
1 J. Friedrich, W. Unger, A. Lippitz, I. Koprinarov, A. Ghode, S. H. Geng and G. Kühn, Composite Interfaces 10(2-3) (2003) 139-171
2 Nathalie Zammatteo et al. Anal. Biochemistry 280 (2000) 143-150
3 W. R. Ashurst, C. Carraro, R. Maboudian, W. Frey Sensors and Actuators A 104 (2003) 213-221
4 G.-Y. Jung et al., Langmuir 21 (2005) 1158-1161.