| Autor: |
Di Natale C; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.; InterdisciplinaryResearch Centre on Biomaterials (CRIB), Università degli Studi di Napoli 'Federico II', Piazzale Tecchio 80, 80125 Naples, Italy., Battista E; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.; InterdisciplinaryResearch Centre on Biomaterials (CRIB), Università degli Studi di Napoli 'Federico II', Piazzale Tecchio 80, 80125 Naples, Italy., Lettera V; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.; Biopox srl, Viale Maria Bakunin 12, 80125 Naples, Italy., Reddy N; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy., Pitingolo G; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy., Vecchione R; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy., Causa F; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.; InterdisciplinaryResearch Centre on Biomaterials (CRIB), Università degli Studi di Napoli 'Federico II', Piazzale Tecchio 80, 80125 Naples, Italy.; Dipartimento di Ingegneria Chimica del Materiali e della Produzione Industriale (DICMAPI), University 'Federico II', Piazzale Tecchio 80, 80125 Naples, Italy., Netti PA; Center for Advanced Biomaterials for Healthcare@CRIB, Istituto Italiano di Tecnologia (IIT), Largo Barsanti e Matteucci 53, 80125 Naples, Italy.; InterdisciplinaryResearch Centre on Biomaterials (CRIB), Università degli Studi di Napoli 'Federico II', Piazzale Tecchio 80, 80125 Naples, Italy.; Dipartimento di Ingegneria Chimica del Materiali e della Produzione Industriale (DICMAPI), University 'Federico II', Piazzale Tecchio 80, 80125 Naples, Italy. |
| Abstrakt: |
The development of assays for protein biomarkers in complex matrices is a demanding task that still needs implementation of new approaches. Antibodies as capture agents have been largely used in bioassays but their low stability, low-efficiency production, and cross-reactivity in multiplex approaches impairs their larger applications. Instead, synthetic peptides, even with higher stability and easily adapted amino acid sequences, still remain largely unexplored in this field. Here, we provide a proof-of-concept of a microfluidic device for direct detection of biomarker overexpression. The multichannel microfluidic polydimethylsiloxane (PDMS) device was first derivatized with PAA (poly(acrylic acid)) solution. CRP-1, VEGF-114, and ΦG6 peptides were preliminarily tested to respectively bind the biomarkers, C-reactive protein (CRP), vascular endothelial growth factor (VEGF), and tumor necrosis factor-alpha (TNF-α). Each PDMS microchannel was then respectively bioconjugated with a specific peptide (CRP-1, VEGF-114, or ΦG6) to specifically capture CRP, VEGF, and TNF-α. With such microdevices, a fluorescence bioassay has been set up with sensitivity in the nanomolar range, both in buffered solution and in human serum. The proposed multiplex assay worked with a low amount of sample (25 μL) and detected biomarker overexpression (above nM concentration), representing a noninvasive and inexpensive screening platform. |
