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A biosensor chip is developed for the detection of a protein biomarker of endocrine disrupting compounds, vitellogenin (Vg) in aquatic environment. The sensor chip is fabricated by immobilizing anti-Vg antibody on 4-Aminothiophenol (4-ATP) coated nanosculptured thin films (nSTFs) of silver on Si substrates. The biosensor is based on the SERS of 4-ATP, enhanced by the Ag nSTFs. Before the fabrication of the sensor, the performance of the enhancement is optimized with respect to the porosity of nSTFs. Further, the biosensor is developed on the nSTF with optimized enhancement. The SERS signals are recorded from the sensor chip for varying concentrations of Vg. A control experiment is performed on another similar protein Fetuin to confirm the specificity of the sensor. The repeatability and reusability of the sensor, along with its shelf life are also checked. The limit of detection of the sensor is found to be 5 pg mL −1 of Vg in PBS within our experimental window. Apart from high sensitivity, specificity and reusability, the present sensor provides additional advantages of miniaturization, requirement of very small volumes of the analyte solution (15 μL) and fast response as compared to conventional techniques e.g., ELISA, as its response time is less than 3 minutes.

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A nanobiosensor chip, utilizing surface enhanced Raman spectroscopy (SERS) on nanosculptured thin films (nSTFs) of silver, was shown to detect Escherichia coli (E. coli) bacteria down to the concentration level of a single bacterium. The sensor utilizes highly enhanced plasmonic nSTFs of silver on a silicon platform for the enhancement of Raman bands as checked with adsorbed 4-aminothiophenol molecules. T-4 bacteriophages were immobilized on the aforementioned surface of the chip for the specific capture of target E. coli bacteria. To demonstrate that no significant non-specific immobilization of other bacteria occurs, three different, additional bacterial strains, Chromobacterium violaceum, Paracoccus denitrificans and Pseudomonas aeruginosa were used. Furthermore, experiments performed on an additional strain of E. coli to address the specificity and reusability of the sensor showed that the sensor operates for different strains of E. coli and is reusable. Time resolved phase contrast microscopy of the E. coli-T4 bacteriophage chip was performed to study its interaction with bacteria over time. Results showed that the present sensor performs a fast, accurate and stable detection of E. coli with ultra-small concentrations of bacteria down to the level of a single bacterium in 10 μl volume of the sample.