Biosensors are key elements in the development of in-vitro diagnostic systems and they permit the fabrication of miniaturized biomedical devices which enable a faster and more reliable diagnostic of various diseases. These will facilitate a fast analysis of analytes present in blood, urine, membranes or saliva and will report the concentration of certain components (biomarkers, drugs, antibiotics, viruses, etc.). Biosensors can also be applied in other fields such as, food, agriculture, monitoring of industrial processes and environmental control. Therefore, the development of low cost, small, sensitive and easy to use biosensors will facilitate the control of diseases, the detection of pollutants and toxins, or the improvement in the production and control of industrial processes. Biosensors are often defined as compact analytical systems, containing a biological recognition element (DNA, antibodies, etc.) that detects a certain analyte (glucose, antigen, etc.) and a transducer that accomplishes the translation of the binding event into a measurable signal for analyte quantification. _x000D_ In this work, electrochemical biosensors have been developed taking advantage of the miniaturization and integration of Lab on a Chip (LOC) devices and of the improvement in sensitivity provided by the use of micro-nanomaterials that enable the production of advanced electrochemical microsensors._x000D_ Thereby, a portable electrochemical detection system (EDS), which is integrated in a LOC device, has been developed. This EDS, consist of a potentiostat and a microfluidic cartridge with a sensor included in it. Here, the operation of this EDS has been mainly demonstrated for DNA measurements. The results obtained show that the response and sensitivity of the EDS are comparable to those obtained using a commercial potentiostat. Besides, since for DNA measurements, which were performed in solution, the sensor surface is not previously modified, this cartridge could be used for the detection of other analytes of clinical interest, as it is demonstrated for uric acid detection._x000D_ With respect to the improvement in sensitivity obtained in the electrochemical measurements, this has been achieved by carbon nanotube (CNT) wiring of magnetic particles (MP) on the surface of the sensor’s working electrode. This enables, on the one hand, the simple and fast acquisition of nanostructured electrode surfaces of improved performance and, on the other hand, the efficient sensing of the MP surface after analyte capture and concentration from the solution. As it is shown in this work, this nanostructuration procedure is applicable to either screen printed or thin film electrodes. Furthermore, CNT wiring of the MP surface has been successfully applied to both the direct biosensing of electroactive and non electroactive molecules, and the enhanced detection of enzymatic markers and targets. In the first case, dopamine detection has been accomplished, obtaining a limit of detection (LOD) low enough to permit the estimation of endogenous dopamine concentration in urine. In the case of the detection of non electroactive analytes (such as antibodies and surfactant), non enzymatic markers have been used (Cd nanocrystals and ferrocene). For these detections, a sandwich assay and a novel detection assay format based on “surface-shadowing” have been exploited, respectively. This latter novel methodology consists in the capture of the analyte, which then interferes in subsequent CNT wiring and electrochemical detection of an electroactive marker that has been previously incorporated to the MP surface. At last, the biosensing of enzymatic targets (myeloperoxidase) has been performed. The LOD obtained in this case was lower than the numbers obtained without using CNT wiring and was appropriate for the study of human serum samples._x000D_ The combination of nanostructured electrode surfaces and a portable EDS has resulted in the development of a LOC device with nanostructured electrodes, applicable to sensitive, fast, portable and potentially automatizable analyte detection.
| Date of Award | 15 Dec 2014 |
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| Original language | Spanish |
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| Supervisor | Eva Baldrich Rubio (Director) & Fernando Martínez Rodríguez (Director) |
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