Enzyme-linked, aptamer-based, competitive biolayer interferometry biosensor for palytoxin.

In this study, we coupled biolayer interferometry (BLI) with competitive binding assay through an enzyme-linked aptamer and developed a real-time, ultra-sensitive, rapid quantitative method for detection of the marine biotoxin palytoxin. Horseradish peroxidase-labeled aptamers were used as biorecognition receptors to competitively bind with palytoxin, which was immobilized on the biosensor surface. The palytoxin: horseradish peroxidase-aptamer complex was then submerged in a 3,3'-diaminobenzidine solution, which resulted in formation of a precipitated polymeric product directly on the biosensor surface and a large change in the optical thickness of the biosensor layer. This change could obviously shift the interference pattern and generate a response profile on the BLI biosensor. The biosensor showed a broad linear range for palytoxin (200-700pg/mL) with a low detection limit (0.04pg/mL). Moreover, the biosensor was applied to the detection of palytoxin in spiked extracts and showed a high degree of selectivity for palytoxin, good reproducibility, and stability. This enzyme-linked, aptamer-based, competitive BLI biosensor offers a promising method for rapid and sensitive detection of palytoxin and other analytes.

[1]  P. Katikou,et al.  First episode of shellfish contamination by palytoxin-like compounds from Ostreopsis species (Aegean Sea, Greece). , 2008, Toxicon : official journal of the International Society on Toxinology.

[2]  P. Katikou,et al.  Occurrence of palytoxin-group toxins in seafood and future strategies to complement the present state of the art. , 2011, Toxicon : official journal of the International Society on Toxinology.

[3]  L. Botana,et al.  Detection of palytoxin-like compounds by a flow cytometry-based immunoassay supported by functional and analytical methods. , 2016, Analytica chimica acta.

[4]  G. Del Favero,et al.  Case definitions for human poisonings postulated to palytoxins exposure. , 2011, Toxicon : official journal of the International Society on Toxinology.

[5]  Danfeng Yao,et al.  Label-free detection of biomolecular interactions using BioLayer interferometry for kinetic characterization. , 2009, Combinatorial chemistry & high throughput screening.

[6]  J. Franco,et al.  Analysis of palytoxin-like in Ostreopsis cultures by liquid chromatography with precolumn derivatization and fluorescence detection , 2006 .

[7]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[8]  C. Ménard,et al.  Quantitative determination of paralytic shellfish poisoning toxins in shellfish using prechromatographic oxidation and liquid chromatography with fluorescence detection: interlaboratory study. , 2004, Journal of AOAC International.

[9]  G. Palleschi,et al.  Development of a haemolytic–enzymatic assay with mediated amperometric detection for palytoxin analysis: application to mussels , 2014, Analytical and Bioanalytical Chemistry.

[10]  Lianghua Wang,et al.  Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application. , 2016, Biosensors & bioelectronics.

[11]  S. Gasparini,et al.  Interactions between Scientists, Managers and Policy Makers in the Framework of the French MediOs Project on Ostreopsis (2008–2010) , 2012 .

[12]  Maurizio Prato,et al.  Highly sensitive electrochemiluminescent nanobiosensor for the detection of palytoxin. , 2012, ACS nano.

[13]  M. C. Louzao,et al.  Marine toxins and the cytoskeleton: a new view of palytoxin toxicity , 2008, The FEBS journal.

[14]  Hao Zhou,et al.  Study of the binding mechanism between aptamer GO18-T-d and gonyautoxin 1/4 by molecular simulation. , 2016, Physical chemistry chemical physics : PCCP.

[15]  K. Kuča,et al.  Toxic potential of palytoxin , 2015, Journal of Huazhong University of Science and Technology [Medical Sciences].

[16]  M. Vieytes,et al.  Palytoxin detection and quantification using the fluorescence polarization technique. , 2012, Analytical biochemistry.

[17]  S. Golubić,et al.  First Evidence of Palytoxin and 42-Hydroxy-palytoxin in the Marine Cyanobacterium Trichodesmium , 2011, Marine drugs.

[18]  Jonathan Deeds,et al.  Human risk associated with palytoxin exposure. , 2010, Toxicon : official journal of the International Society on Toxinology.

[19]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[20]  C. Dell’Aversano,et al.  LC-MS of palytoxin and its analogues: State of the art and future perspectives. , 2011, Toxicon : official journal of the International Society on Toxinology.

[21]  M. Vieytes,et al.  Surface Plasmon Resonance Biosensor Method for Palytoxin Detection Based on Na+,K+-ATPase Affinity , 2013, Toxins.

[22]  S. Jayasena.  Aptamers: an emerging class of molecules that rival antibodies in diagnostics. , 1999, Clinical chemistry.

[23]  J. Franco,et al.  Palytoxins: biological and chemical determination. , 2011, Toxicon : official journal of the International Society on Toxinology.

[24]  A. Penna,et al.  Investigation of toxin profile of Mediterranean and Atlantic strains of Ostreopsis cf. siamensis (Dinophyceae) by liquid chromatography–high resolution mass spectrometry , 2013 .

[25]  M. Murado,et al.  Mouse bioassay for palytoxin. Specific symptoms and dose-response against dose-death time relationships. , 2008, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[26]  Y. Chisti,et al.  Bioactives from microalgal dinoflagellates. , 2012, Biotechnology advances.

[27]  Richard E. Moore,et al.  Palytoxin: A New Marine Toxin from a Coelenterate , 1971, Science.

[28]  S. Kumaraswamy,et al.  Label-free kinetic analysis of an antibody-antigen interaction using biolayer interferometry. , 2015, Methods in molecular biology.

[29]  E. Ferapontova,et al.  An RNA aptamer-based electrochemical biosensor for detection of theophylline in serum. , 2008, Journal of the American Chemical Society.

[30]  Weihong Tan,et al.  Aptamers from cell-based selection for bioanalytical applications. , 2013, Chemical reviews.