Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4
Here, we present the results of our the electrochemical aptasensing strategy for retinol binding protein-4 (RBP-4) detection based on a thiolated aptamer against RBP-4 and 6-mercaptohexanol (MCH) directly immobilized on a gold electrode surface. The most important parameters affecting the magnitude...
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| Format: | Article |
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MDPI AG
2024-02-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/14/2/101 |
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| author | Kamila Malecka-Baturo Paulina Żółtowska Agnieszka Jackowska Katarzyna Kurzątkowska-Adaszyńska Iwona Grabowska |
| author_facet | Kamila Malecka-Baturo Paulina Żółtowska Agnieszka Jackowska Katarzyna Kurzątkowska-Adaszyńska Iwona Grabowska |
| author_sort | Kamila Malecka-Baturo |
| collection | DOAJ |
| description | Here, we present the results of our the electrochemical aptasensing strategy for retinol binding protein-4 (RBP-4) detection based on a thiolated aptamer against RBP-4 and 6-mercaptohexanol (MCH) directly immobilized on a gold electrode surface. The most important parameters affecting the magnitude of the analytical signal generated were optimized: (i) the presence of magnesium ions in the immobilization and measurement buffer, (ii) the concentration of aptamer in the immobilization solution and (iii) its folding procedure. In this work, a systematic assessment of the electrochemical parameters related to the optimization of the sensing layer of the aptasensor was carried out (electron transfer coefficients <i>(α),</i> electron transfer rate constants (<i>k</i><sup>0</sup>) and surface coverage of the thiolated aptamer probe (<i>Γ<sub>Apt</sub></i>)). Then, under the optimized conditions, the analytical response towards RBP-4 protein, in the presence of an Fe(CN)<sub>6</sub><sup>3−/4−</sup> redox couple in the supporting solution was assessed. The proposed electrochemical strategy allowed for RBP-4 detection in the concentration range between 100 and 1000 ng/mL with a limit of detection equal to 44 ng/mL based on electrochemical impedance spectroscopy (EIS). The specificity studies against other diabetes biomarkers, including vaspin and adiponectin, proved the selectivity of the proposed platform. These preliminary results will be used in the next step to miniaturize and test the sensor in real samples. |
| first_indexed | 2024-03-07T22:40:10Z |
| format | Article |
| id | doaj.art-f0792425194247a7ab15a20c6ddcc5ea |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-07T22:40:10Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-f0792425194247a7ab15a20c6ddcc5ea2024-02-23T15:09:35ZengMDPI AGBiosensors2079-63742024-02-0114210110.3390/bios14020101Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4Kamila Malecka-Baturo0Paulina Żółtowska1Agnieszka Jackowska2Katarzyna Kurzątkowska-Adaszyńska3Iwona Grabowska4Institute of Animal Reproduction and Food Research Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, PolandDepartment of Chemistry, University of Warmia and Mazury, Plac Łódzki 4, 10-721 Olsztyn, PolandDepartment of Chemistry, University of Warmia and Mazury, Plac Łódzki 4, 10-721 Olsztyn, PolandInstitute of Animal Reproduction and Food Research Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, PolandInstitute of Animal Reproduction and Food Research Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, PolandHere, we present the results of our the electrochemical aptasensing strategy for retinol binding protein-4 (RBP-4) detection based on a thiolated aptamer against RBP-4 and 6-mercaptohexanol (MCH) directly immobilized on a gold electrode surface. The most important parameters affecting the magnitude of the analytical signal generated were optimized: (i) the presence of magnesium ions in the immobilization and measurement buffer, (ii) the concentration of aptamer in the immobilization solution and (iii) its folding procedure. In this work, a systematic assessment of the electrochemical parameters related to the optimization of the sensing layer of the aptasensor was carried out (electron transfer coefficients <i>(α),</i> electron transfer rate constants (<i>k</i><sup>0</sup>) and surface coverage of the thiolated aptamer probe (<i>Γ<sub>Apt</sub></i>)). Then, under the optimized conditions, the analytical response towards RBP-4 protein, in the presence of an Fe(CN)<sub>6</sub><sup>3−/4−</sup> redox couple in the supporting solution was assessed. The proposed electrochemical strategy allowed for RBP-4 detection in the concentration range between 100 and 1000 ng/mL with a limit of detection equal to 44 ng/mL based on electrochemical impedance spectroscopy (EIS). The specificity studies against other diabetes biomarkers, including vaspin and adiponectin, proved the selectivity of the proposed platform. These preliminary results will be used in the next step to miniaturize and test the sensor in real samples.https://www.mdpi.com/2079-6374/14/2/101electrochemical aptasensoroptimization processretinol binding protein-4RBP-4 |
| spellingShingle | Kamila Malecka-Baturo Paulina Żółtowska Agnieszka Jackowska Katarzyna Kurzątkowska-Adaszyńska Iwona Grabowska Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4 Biosensors electrochemical aptasensor optimization process retinol binding protein-4 RBP-4 |
| title | Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4 |
| title_full | Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4 |
| title_fullStr | Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4 |
| title_full_unstemmed | Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4 |
| title_short | Electrochemical Aptasensing Platform for the Detection of Retinol Binding Protein-4 |
| title_sort | electrochemical aptasensing platform for the detection of retinol binding protein 4 |
| topic | electrochemical aptasensor optimization process retinol binding protein-4 RBP-4 |
| url | https://www.mdpi.com/2079-6374/14/2/101 |
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