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An array-based nanosensor for detecting cellular responses in macrophages induced by femtomolar levels of pesticides.

ABSTRACT: Environmental agents can induce cellular responses at concentrations far below the limits of detection for current viability and biomarker-based cell sensing platforms. Hypothesis-free cell sensor platforms can be engineered to maximize sensitivity to phenotypic changes, providing a tool for lowering the threshold for detecting cellular changes. Pesticides are one of the most prevalent sources of chemical exposure due to their use in food and agriculture fields. We report here a FRET-based nanosensor array engineered to maximize responses to changes at cell surfaces after pesticide exposure. This sensor array robustly detected macrophage responses to femtomolar concentrations of common pesticides-orders of magnitude lower concentrations than traditional toxicological and biomarker-based strategies. Significantly, this platform was able to classify these responses by pesticide class, demonstrating the ability to distinguish between changes induced by these different agents. Taken together, hypothesis-free cell surface sensing is a promising tool for detecting the effects of ultra-trace environmental chemicals on human health, as well as detecting threshold responses for use in drug discovery and diagnostics.

SUBMITTER: Jiang M

PROVIDER: S-EPMC10587896 | biostudies-literature | 2022 Feb

REPOSITORIES: biostudies-literature

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An array-based nanosensor for detecting cellular responses in macrophages induced by femtomolar levels of pesticides.

Jiang Mingdi M Chattopadhyay Aritra Nath AN Geng Yingying Y Rotello Vincent M VM

Chemical communications (Cambridge, England) 20220224 17

Environmental agents can induce cellular responses at concentrations far below the limits of detection for current viability and biomarker-based cell sensing platforms. Hypothesis-free cell sensor platforms can be engineered to maximize sensitivity to phenotypic changes, providing a tool for lowering the threshold for detecting cellular changes. Pesticides are one of the most prevalent sources of chemical exposure due to their use in food and agriculture fields. We report here a FRET-based nanos ...[more]

PMID: 35141736

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Project description:In the search of new robust and environmental-friendly analytical methods able to answer quantitative issues in pharmacology, we explore liquid chromatography (LC) associated with elemental mass spectrometry (ICP-MS) to monitor peptides in such complex biological matrices. The novelty is to use mass spectrometry to replace radiolabelling and radioactivity measurements, which represent up-to now the gold standard to measure organic compound concentrations in life science. As a proof of concept, we choose the vasopressin (AVP)/V1A receptor system for model pharmacological assays. The capacity of ICP-MS to provide highly sensitive quantitation of metallic and hetero elements, whatever the sample medium, prompted us to investigate this technique in combination with appropriate labelling of the peptide of interest. Selenium, that is scarcely present in biological media, was selected as a good compromise between ICP-MS response, covalent tagging ability using conventional sulfur chemistry and peptide detection specificity. Applying selenium monitoring by elemental mass spectrometry in pharmacology is challenging due to the very high salt content and organic material complexity of the samples that produces polyatomic aggregates and thus potentially mass interferences with selenium detection. Hyphenation with a chromatographic separation was found compulsory. Noteworthy, we aimed to develop a straightforward quantitative protocol that can be performed in any laboratory equipped with a standard macrobore LC-ICP-MS system, in order to avoid time-consuming sample treatment or special implementation of instrumental set-up, while allowing efficient suppression of all mass interferences to reach the targeted sensitivity. Significantly, a quantification limit of 57 ng Se L-1 (72 femtomoles of injected Se) was achieved, the samples issued from the pharmacological assays being directly introduced into the LC-ICP-MS system. The established method was successfully validated and applied to the measurement of the vasopressin ligand affinity for its V1A receptor through the determination of the dissociation constant (Kd) which was compared to the one recorded with conventional radioactivity assays.

Dataset Information

An array-based nanosensor for detecting cellular responses in macrophages induced by femtomolar levels of pesticides.

Publications

An array-based nanosensor for detecting cellular responses in macrophages induced by femtomolar levels of pesticides.

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