Abstract
The increasing demand for easily available and low-cost diagnostics has fuelled the development of aptasensors as platforms for rapid, sensitive, and point-of-care testing of target analytes. Recently, gold nanoparticle (AuNP)-based aptasensors have attracted wide recognition owing to their color transition properties which allow real-time rapid sensing of targets. In this study, we utilized the color transition property of aptamer-functionalized AuNPs to detect and quantify estrogen receptor alpha (ERα), a key biomarker protein in breast cancer. We found that the coating of AuNPs with unmodified ERα-RNA aptamer (GGGGUCAAGGUGACCCC) makes them resistant to salt-induced aggregation. However, addition of ERα to the aptamer-protected AuNPs results in their spontaneous aggregation as evident from a color transition from wine red to deep blue. On the basis of this, we developed an ERα aptasensor, with limits of detection and quantification of 0.64 and 2.16 ng/mL, respectively; the aptasensor can efficiently detect and quantify ERα in a working range of 10 ng/mL–5μg/mL protein. Validation of the aptasensor on cellular extracts of ERα-positive MCF-7 and ERα-deficient MDA-MB-231 breast cancer cells showed a target-selective response in ERα-positive samples but not in cellular extracts of ERα-deficient breast cancer cells. Further, the small size and simple fabrication chemistry of aptamers provide an additional benefit to make the ERα aptasensor a potentially useful and cost-effective tool in point-of-care analyses of ERα.
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Acknowledgments
R. A. thanks the Council of Scientific and Industrial Research, Government of India for the award of a senior research fellowship. This work was financially supported by CARDIOMED project no. BSC 0122 of the Council of Scientific and Industrial Research, Government of India, New Delhi, India.
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Ahirwar, R., Nahar, P. Development of a label-free gold nanoparticle-based colorimetric aptasensor for detection of human estrogen receptor alpha. Anal Bioanal Chem 408, 327–332 (2016). https://doi.org/10.1007/s00216-015-9090-7
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DOI: https://doi.org/10.1007/s00216-015-9090-7