A Novel Sensitive Cell-Based Immunoenzymatic Assay for Palytoxin Quantitation in Mussels

Pelin, Marco; Sosa, Silvio; Brovedani, Valentina; Fusco, Laura; Poli, Mark; Tubaro, Aurelia

doi:10.3390/toxins10080329

Cited by 8 publications

(7 citation statements)

References 62 publications

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“…The recorded EC 50 and Emax values demonstrate a wide inter-individual variability in the in vitro sensitivity towards PLTX. This result corroborates our previous in vitro findings using a panel of 9 different cell lines, derived from different tissues and showing a wide range of sensitivity for PLTX binding [44]. However, the present results were obtained using the same cell type (i.e., healthy volunteers' monocytes), therefore avoiding variable sensitivity due to differences in metabolic pathways and/or signal transduction pathways characterizing different cell types, such as in the previous study.…”

Section: Discussionsupporting

confidence: 92%

In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform

Pelin

Stocco

Florio

et al. 2020

IJMS

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The marine polyether palytoxin (PLTX) is one of the most toxic natural compounds, and is involved in human poisonings after oral, inhalation, skin and/or ocular exposure. Epidemiological and molecular evidence suggest different inter-individual sensitivities to its toxic effects, possibly related to genetic-dependent differences in the expression of Na+/K+-ATPase, its molecular target. To identify Na+/K+-ATPase subunits, isoforms correlated with in vitro PLTX cytotoxic potency, sensitivity parameters (EC50: PLTX concentration reducing cell viability by 50%; Emax: maximum effect induced by the highest toxin concentration; 10−7 M) were assessed in 60 healthy donors’ monocytes by the MTT (methylthiazolyl tetrazolium) assay. Sensitivity parameters, not correlated with donors’ demographic variables (gender, age and blood group), demonstrated a high inter-individual variability (median EC50 = 2.7 × 10−10 M, interquartile range: 0.4–13.2 × 10−10 M; median Emax = 92.0%, interquartile range: 87.5–94.4%). Spearman’s analysis showed significant positive correlations between the β2-encoding ATP1B2 gene expression and Emax values (rho = 0.30; p = 0.025) and between Emax and the ATP1B2/ATP1B3 expression ratio (rho = 0.38; p = 0.004), as well as a significant negative correlation between Emax and the ATP1B1/ATP1B2 expression ratio (rho = −0.30; p = 0.026). This toxicogenetic study represents the first approach to define genetic risk factors that may influence the onset of adverse effects in human PLTX poisonings, suggesting that individuals with high gene expression pattern of the Na+/K+-ATPase β2 subunit (alone or as β2/β1 and/or β2/β3 ratio) could be highly sensitive to PLTX toxic effects.

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Section: Discussionsupporting

confidence: 92%

In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform

Pelin

Stocco

Florio

et al. 2020

IJMS

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“…Pelin et al [ 79 ] have taken a different approach in their PLTX assay. Instead of competitive-based inhibition, their assay was designed as a sandwich; PLTX is captured by receptors on immobilized HaCaT cells and then detected with labelled antibodies.…”

Section: Methods For Official Control Testingmentioning

confidence: 99%

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

Dillon

Zaczek-Moczydlowska

Edwards

et al. 2021

Sensors

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In the past twenty years marine biotoxin analysis in routine regulatory monitoring has advanced significantly in Europe (EU) and other regions from the use of the mouse bioassay (MBA) towards the high-end analytical techniques such as high-performance liquid chromatography (HPLC) with tandem mass spectrometry (MS). Previously, acceptance of these advanced methods, in progressing away from the MBA, was hindered by a lack of commercial certified analytical standards for method development and validation. This has now been addressed whereby the availability of a wide range of analytical standards from several companies in the EU, North America and Asia has enhanced the development and validation of methods to the required regulatory standards. However, the cost of the high-end analytical equipment, lengthy procedures and the need for qualified personnel to perform analysis can still be a challenge for routine monitoring laboratories. In developing regions, aquaculture production is increasing and alternative inexpensive Sensitive, Measurable, Accurate and Real-Time (SMART) rapid point-of-site testing (POST) methods suitable for novice end users that can be validated and internationally accepted remain an objective for both regulators and the industry. The range of commercial testing kits on the market for marine toxin analysis remains limited and even more so those meeting the requirements for use in regulatory control. Individual assays include enzyme-linked immunosorbent assays (ELISA) and lateral flow membrane-based immunoassays (LFIA) for EU-regulated toxins, such as okadaic acid (OA) and dinophysistoxins (DTXs), saxitoxin (STX) and its analogues and domoic acid (DA) in the form of three separate tests offering varying costs and benefits for the industry. It can be observed from the literature that not only are developments and improvements ongoing for these assays, but there are also novel assays being developed using upcoming state-of-the-art biosensor technology. This review focuses on both currently available methods and recent advances in innovative methods for marine biotoxin testing and the end-user practicalities that need to be observed. Furthermore, it highlights trends that are influencing assay developments such as multiplexing capabilities and rapid POST, indicating potential detection methods that will shape the future market.

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“…Subsequently, the solid-phase magnetic particles containing the bound toxins were magnetically conducted through the liquid-stationary phase into the detection chamber containing the HRP substrate [66]. Pelin et al (2018) were able to conjugate cell-based assays with immunoenzymatic detection for the quantification of palytoxin. The biotoxin binds with high affinity to several cell lines and could then be detected by the addition of a specific antibody [67].…”

Section: Biochemical Assaysmentioning

confidence: 99%

“…Pelin et al (2018) were able to conjugate cell-based assays with immunoenzymatic detection for the quantification of palytoxin. The biotoxin binds with high affinity to several cell lines and could then be detected by the addition of a specific antibody [67].…”

Section: Biochemical Assaysmentioning

confidence: 99%

“…Sensors 2021, 21, x FOR PEER REVIEW 6 of 21 palytoxin. The biotoxin binds with high affinity to several cell lines and could then be detected by the addition of a specific antibody [67]. Despite advantages in simplicity, sensitivity, and selectivity, immunoassays are still associated with high laboratory and antibody preparation costs, antibody instability, and the possibility of false-positive or false-negative results [68].…”

Section: Optical Biosensors For Detecting Aquatic Biotoxinsmentioning

confidence: 99%

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Emerging Optical Materials in Sensing and Discovery of Bioactive Compounds

Vaz

Valpradinhos

Frasco

et al. 2021

Sensors

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Optical biosensors are used in numerous applications and analytical fields. Advances in these sensor platforms offer high sensitivity, selectivity, miniaturization, and real-time analysis, among many other advantages. Research into bioactive natural products serves both to protect against potentially dangerous toxic compounds and to promote pharmacological innovation in drug discovery, as these compounds have unique chemical compositions that may be characterized by greater safety and efficacy. However, conventional methods for detecting these biomolecules have drawbacks, as they are time-consuming and expensive. As an alternative, optical biosensors offer a faster, simpler, and less expensive means of detecting various biomolecules of clinical interest. In this review, an overview of recent developments in optical biosensors for the detection and monitoring of aquatic biotoxins to prevent public health risks is first provided. In addition, the advantages and applicability of these biosensors in the field of drug discovery, including high-throughput screening, are discussed. The contribution of the investigated technological advances in the timely and sensitive detection of biotoxins while deciphering the pathways to discover bioactive compounds with great health-promoting prospects is envisaged to meet the increasing demands of healthcare systems.

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A Novel Sensitive Cell-Based Immunoenzymatic Assay for Palytoxin Quantitation in Mussels

Cited by 8 publications

References 62 publications

In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform

In Vitro Cell Sensitivity to Palytoxin Correlates with High Gene Expression of the Na+/K+-ATPase β2 Subunit Isoform

Current Trends and Challenges for Rapid SMART Diagnostics at Point-of-Site Testing for Marine Toxins

Emerging Optical Materials in Sensing and Discovery of Bioactive Compounds

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