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EnglishImmunosensors are one of the most common platform used in clinical laboratories, in particular the class based on Enzyme Linked Fluorescent Assays (ELFA) takes advantage of the amplification step of the enzyme, usually the alkaline phosphatase, that catalyzes the hydrolysis of a fluorescent substrate leading it to fluoresce. Anyway, they suffer in sensitivity if compared to molecular diagnostic or more modern in vitro diagnostic devices. In our work, a simple and effective mechanism to enhance the fluorescent signal, and hence the sensitivity of the system, is presented. It is based on the metal ion displacement principle in which a second fluorophore, in our case Calcein Blue, quenched by a cobalt ion is add to the first one (4-MUP), and, in presence of inorganic phosphate, it will be progressively activated by the inorganic phosphate itself leading to the metal displacement. In this way Calcein Blue, newly free to fluoresce, contributes to global fluorescent signal generated by 4-MU. We have tested our proof of principle on a currently used immunoanalyzer, that is VIDAS® system (bioMérieux, Marcy l'Etoile, France) obtaining a fluorescence enhancement of about 50% for each concentration of hydrolyzed 4-MUP tested.
| Autori interni: | |
| Autori: | Susini, V.; Ienco, A.; Lucia Rossi, V.; Paolicchi, A.; Sanesi, A. |
| Titolo: | Fluorescence enhancement aided by metal ion displacement |
| Presenza coautori internazionali: | no |
| Numero degli autori: | 5 |
| Anno del prodotto: | 2016 |
| Rivista: | BIOSENSORS & BIOELECTRONICS |
| Lingua: | Inglese |
| Volume: | 80 |
| Pagina iniziale: | 237 |
| Pagina finale: | 242 |
| Numero di pagine: | 6 |
| Digital Object Identifier (DOI): | http://dx.doi.org/10.1016/j.bios.2016.01.062 |
| Codice identificativo Pubmed: | 26851581 |
| URL: | https://www.sciencedirect.com/science/article/pii/S0956566316300707 |
| IF: | si |
| Codice identificativo Scopus: | 2-s2.0-84956650891 |
| Codice identificativo ISI: | WOS:000372558500035 |
| Abstract: | Immunosensors are one of the most common platform used in clinical laboratories, in particular th...e class based on Enzyme Linked Fluorescent Assays (ELFA) takes advantage of the amplification step of the enzyme, usually the alkaline phosphatase, that catalyzes the hydrolysis of a fluorescent substrate leading it to fluoresce. Anyway, they suffer in sensitivity if compared to molecular diagnostic or more modern in vitro diagnostic devices. In our work, a simple and effective mechanism to enhance the fluorescent signal, and hence the sensitivity of the system, is presented. It is based on the metal ion displacement principle in which a second fluorophore, in our case Calcein Blue, quenched by a cobalt ion is add to the first one (4-MUP), and, in presence of inorganic phosphate, it will be progressively activated by the inorganic phosphate itself leading to the metal displacement. In this way Calcein Blue, newly free to fluoresce, contributes to global fluorescent signal generated by 4-MU. We have tested our proof of principle on a currently used immunoanalyzer, that is VIDAS® system (bioMérieux, Marcy l'Etoile, France) obtaining a fluorescence enhancement of about 50% for each concentration of hydrolyzed 4-MUP tested. |
| Data di presentazione: | 11-gen-2021 |
| Parole Chiave: | Fluorescence enhancement; Immunosensors; Metal ion displacement; Cobalt; Fluoresceins; Fluorescent Dyes; Humans; Hydrolysis; Biosensing Techniques; Enzyme Assays; Reagent Kits, Diagnostic |
| Appare nelle tipologie: | 1.1 Articolo in rivista |
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