Publicação
Dual-aptasensor array for osteopontin detection: optimization of aptamers immobilization conditions
| Resumo: | Cancer diseases are associated with the presence of several protein biomarkers. Aptasensor arrays may allow early multiple-detection of these biomarkers which can make significant improvements in the lives of cancer patients. Clinical studies suggest that osteopontin, an overexpressed protein by tumor cells, may be used as a diagnostic biomarker for various cancers. The aim of the present work was to establish the optimal experimental conditions that allow enhancing the performance of an electrochemical dual-aptasensor array for the detection of osteopontin. The aptamer concentration, time and temperature of immobilization into the dual-screen printed gold electrodes (Dual-SPGE) as well as the aptamer-protein interaction time were evaluated using a 2k factorial experimental design. DNA and/or RNA aptamers were immobilized on the Dual-SPGE via streptavidin-biotin interaction and the evaluation of the best experimental conditions was carried out by cyclic and square wave voltammetry using a ferro/ferricyanide solution ([Fe(CN)6]3−/4−) as a redox probe. Statistical significant models for both DNA and RNA aptamers were established. Temperature and aptamer concentration were found to be the most significant parameters. Also, the results pointed out that interaction effects between the four parameters were usually statistically significant, showing that there was a dependency between aptamer concentration-aptamer immobilization time and between temperature-aptamer immobilization time. The optimum experimental conditions found for enhancing the performance of the dual-aptasensor array were 4°C, 0.5 μM of aptamer concentration, 20 min of aptamer incubation and 30 min of aptamer-osteopontin interaction. |
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| Autores principais: | Ezziddine, Maha |
| Ano: | 2017 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Instituto Politécnico de Bragança |
| Idioma: | inglês |
| Origem: | Biblioteca Digital do IPB |
| Resumo: | Cancer diseases are associated with the presence of several protein biomarkers. Aptasensor arrays may allow early multiple-detection of these biomarkers which can make significant improvements in the lives of cancer patients. Clinical studies suggest that osteopontin, an overexpressed protein by tumor cells, may be used as a diagnostic biomarker for various cancers. The aim of the present work was to establish the optimal experimental conditions that allow enhancing the performance of an electrochemical dual-aptasensor array for the detection of osteopontin. The aptamer concentration, time and temperature of immobilization into the dual-screen printed gold electrodes (Dual-SPGE) as well as the aptamer-protein interaction time were evaluated using a 2k factorial experimental design. DNA and/or RNA aptamers were immobilized on the Dual-SPGE via streptavidin-biotin interaction and the evaluation of the best experimental conditions was carried out by cyclic and square wave voltammetry using a ferro/ferricyanide solution ([Fe(CN)6]3−/4−) as a redox probe. Statistical significant models for both DNA and RNA aptamers were established. Temperature and aptamer concentration were found to be the most significant parameters. Also, the results pointed out that interaction effects between the four parameters were usually statistically significant, showing that there was a dependency between aptamer concentration-aptamer immobilization time and between temperature-aptamer immobilization time. The optimum experimental conditions found for enhancing the performance of the dual-aptasensor array were 4°C, 0.5 μM of aptamer concentration, 20 min of aptamer incubation and 30 min of aptamer-osteopontin interaction. |
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