http://www.cnr.it/ontology/cnr/individuo/prodotto/ID286807

OncomiR detection in circulating body fluids: A PDMS microdevice perspective (Articolo in rivista)

Type

Articolo in rivista (Classe)
Prodotto della ricerca (Classe)

Label

OncomiR detection in circulating body fluids: A PDMS microdevice perspective (Articolo in rivista) (literal)

Anno

2014-01-01T00:00:00+01:00 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi

10.1039/c4lc00630e (literal)

Alternative label

Potrich C.; Vaghi V.; Lunelli L.; Pasquardini L.; Santini G.C.; Ottone C.; Quaglio M.; Cocuzza M.; Pirri C.F.; Ferracin M.; Negrini M.; Tiberio P.; De Sanctis V.; Bertorelli R.; Pederzolli C. (2014)
OncomiR detection in circulating body fluids: A PDMS microdevice perspective
in Lab on a chip (Online); Royal Society of Chemistry, Cambridge (Regno Unito)
(literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

Potrich C.; Vaghi V.; Lunelli L.; Pasquardini L.; Santini G.C.; Ottone C.; Quaglio M.; Cocuzza M.; Pirri C.F.; Ferracin M.; Negrini M.; Tiberio P.; De Sanctis V.; Bertorelli R.; Pederzolli C. (literal)

Pagina inizio

4067 (literal)

Pagina fine

4075 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#url

http://www.scopus.com/inward/record.url?eid=2-s2.0-84907855169&partnerID=q2rCbXpz (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

14 (literal)

Rivista

Lab on a chip (Rivista)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo

20 (literal)

Note

ISI Web of Science (WOS) (literal)
Scopu (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

Fondazione Bruno Kessler-LaBSSAH, Via Sommarive 18, Trento, Italy; CNR-Institute of Biophysics, Unit at Trento, Via alla Cascata 56, Povo, Trento, 38123, Italy; Center for Advanced Biomaterials for Healthcare, Istituto Italiano di Tecnologia, Largo Barsanti e Matteucci 53, Napoli, 80125, Italy; Center for Space Human Robotics Polito, Istituto Italiano di Tecnologia, C. so Trento 21, Torino, 10129, Italy; Department of Applied Science and Technology, Politecnico di Torino, C. so Duca degli Abruzzi 24, Torino, 10129, Italy; CNR-IMEM, Parco Area delle Scienze 37a, Parma, 43124, Italy; Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, Ferrara, Italy; Unit of Biomarkers, Department of Experimental Oncology and Molecular Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy; NGS Facility-LaBSSAH, Centre for Integrative Biology, University of Trento, Italy (literal)

Titolo

OncomiR detection in circulating body fluids: A PDMS microdevice perspective (literal)

Abstract

There is an increasing interest in circulating microRNAs (miRNAs) as potential minimally invasive diagnostic biomarkers in oncology. Considerable efforts are being made in the development of lab-on-a-chip devices for biomedical applications to purify and detect miRNAs from biological fluids. Here, we report the development of an innovative polydimethylsiloxane (PDMS)-based parallel device whose internal surface can opportunely be functionalized with positively charged 3-aminopropyltriethoxysilane (APTES) alone or mixed with two different neutral poly(ethylene glycol) silanes (PEG-s). The differently functionalized internal surfaces of the PDMS chip were characterized with s-SDTB (sulfosuccinimidyl-4-o-(4,4-dimethoxytrityl) butyrate) and the portion of the surface able to adsorb a synthetic fluorescently labeled miRNA was determined. Interestingly, the adsorbed miRNA (both synthetic and cell supernatant-derived) was found mainly on the bottom surface of the chip and could be reverse transcribed into cDNA directly on the same PDMS chip used for its purification, saving hours with respect to the use of standard purification kits. We identified 0.1% APTES/0.9% PEG-silane as the most efficient PDMS functionalization to capture both synthetic and extracellular miRNA. Moreover, the amount of captured miRNA was increased by treating the cell supernatant with a commercially available lysis buffer for RNA extraction. We assessed that the available miRNA binding sites on the functionalized surface were efficiently saturated with only one incubation, shortening the time and greatly simplifying the protocol for miRNA purification from biological samples. Finally, the extracellular miRNA purification efficiency of the PDMS functionalized multichip determined via real-time quantitative polymerase chain reaction (RT-qPCR) was confirmed by droplet digital PCR (ddPCR) quantification. This work shows an innovative, rapid and easy to use microdevice for the purification and reverse transcription of circulating miRNAs, approaching the realization of diagnostic and prognostic oncomiR-based assays. This journal is (literal)

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