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Probing high-affinity aptamer binding region and development of aptasensor platform for the detection of cylindrospermopsin

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Abstract

Cylindrospermopsin (CYN) is one of the most concerning cyanotoxins due to its potential toxicity and spreading to various environments including drinking water. CYN has potential interferences with human and animal metabolic pathways, which influence the functions of organs including liver, kidneys, lungs, etc. CYN is involved in the inhibition of protein synthesis and detachment of ribosomes from the endoplasmic reticulum membrane. It also interacts with soluble proteins, which are associated with protein translations. It is believed that cytochrome 450 is responsible for the rapid toxicity of CYN. Researchers are urged to develop a high-throughput screening method for the detection of CYN in water. Construction of low cost, rapid, and sensitive analytical methods for the detection of CYN is challenging. Here, we used graphene oxide (GO) as the fluorescence sensing platform for probing the high affinity of the short aptamer derived from the wild-type long aptamer-CYN sensing. The biosensor construction involved two steps: first, quenching the fluorescence of fluorescent-labelled truncated aptamer using GO as a quencher and, second, fluorescence recovery in the presence of CYN by competitive binding between the target and GO. One of the truncate aptamers has a 12-fold higher affinity and enhances sensitivity compared to the long aptamer sequence. The limit of detection of the high affinity truncated aptamer is 17 pM which is 6-fold lower than the long aptamer (100 pM). The sensor specifically detects CYN in the presence of other potential interfering toxins. The performance of the sensor was validated using CYN spiked tap water with very good recovery percentage. A rapid and highly sensitive detection of CYN from water resources has been achieved using this method.

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Acknowledgements

The authors would like to acknowledge the funding from the research office at Alfaisal University.

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Author notes

  1. Raja Chinnappan and Razan AlZabn contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, Alfaisal University, Al Zahrawi Street, Al Maather, Al Takhassusi Road, Riyadh, 11533, Saudi Arabia

    Raja Chinnappan, Razan AlZabn, Amjad K. Fataftah & Mohammed Zourob

  2. Department of Pharmacology and Toxicology, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia

    Ali Alhoshani

  3. King Faisal Specialist Hospital and Research Center, Al Zahrawi Street, Al Maather, Riyadh, 12713, Saudi Arabia

    Mohammed Zourob

Authors
  1. Raja Chinnappan
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  2. Razan AlZabn
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  3. Amjad K. Fataftah
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  4. Ali Alhoshani
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  5. Mohammed Zourob
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Correspondence to Mohammed Zourob.

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Chinnappan, R., AlZabn, R., Fataftah, A.K. et al. Probing high-affinity aptamer binding region and development of aptasensor platform for the detection of cylindrospermopsin. Anal Bioanal Chem 412, 4691–4701 (2020). https://doi.org/10.1007/s00216-020-02723-4

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  • DOI: https://doi.org/10.1007/s00216-020-02723-4

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