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A novel fluorescent biosensor based on affinity-enhanced aptamer-peptide conjugate for sensitive detection of lead(II) in aquatic products

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Abstract

Lead contamination is a major concern in food safety and, as such, many lead detection methods have been developed, especially aptamer-based biosensors. However, the sensitivity and environmental tolerance of these sensors require improvement. A combination of different types of recognition elements is an effective way to improve the detection sensitivity and environmental tolerance of biosensors. Here, we provide a novel recognition element, an aptamer-peptide conjugate (APC), to achieve enhanced affinity of Pb2+. The APC was synthesized from Pb2+ aptamers and peptides through clicking chemistry. The binding performance and environmental tolerance of APC with Pb2+ was studied through isothermal titration calorimetry (ITC); the binding constant (Ka) was 1.76*106 M−1, indicating that the APC’s affinity was increased by 62.96% and 802.56% compared with the aptamers and peptides, respectively. Besides, APC demonstrated better anti-interference (K+) than aptamer and peptide. Through the molecular dynamics (MD) simulation, we found that more binding sites and stronger binding energy between APC with Pb2+are the reasons for higher affinity between APC with Pb2+. Finally, a carboxyfluorescein (FAM)-labeled APC fluorescent probe was synthesized and a fluorescent detection method for Pb2+ was established. The limit of detection of the FAM-APC probe was calculated to be 12.45 nM. This detection method was also applied to the swimming crab and showed great potential in real food matrix detection.

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Funding

National Natural Science Foundation of China (32272421) and National Natural Science Foundation of China (32072312). This work was partly supported by the Science and Technology Innovation Action Plan of Shanghai (20392001600).

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  1. School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai Engineering Research Center for Food Rapid Detection, Shanghai, 200093, China

    Kaimin Peng, ** Wang

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  1. Kaimin Peng
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Peng, K., Liu, X., Yuan, H. et al. A novel fluorescent biosensor based on affinity-enhanced aptamer-peptide conjugate for sensitive detection of lead(II) in aquatic products. Anal Bioanal Chem 415, 3463–3474 (2023). https://doi.org/10.1007/s00216-023-04735-2

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