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Biomedical Microdevices

, 20:60 | Cite as

A magnetic beads-based portable flow cytometry immunosensor for in-situ detection of marine biotoxin

  • Yuxiang Pan
  • Xinwei Wei
  • Tao Liang
  • Jie Zhou
  • Hao Wan
  • Ning Hu
  • Ping Wang
Article

Abstract

Okadaic acid (OA), a representative diarrhetic shellfish poisoning toxin, mainly produced by toxigenic dinoflagellates, has significant hazard to public health. Traditional methods for detection of OA can not give the consideration to the need of rapid, high sensitive, quantitative and in-situ detection at the same time. Herein, a new effective detection method of OA was developed based on fluorescence immunosensor and flow cytometry (FCM). In this assay, Streptavidin-coated magnetic beads (MBs) were used as the supporter to immobilize the biotinylated OA. Modified MBs competed with the free OA in the sample solution to bind with the anti-OA monoclonal antibody (OA-MAb). The R-phycoerythrin (R-PE) dye labeled IgG was served as a secondary antibody to perform fluorescence detection. A portable flow cytometry was applied for the in-situ fluorescence quantification. The results showed that the OA concentration was inversely proportional to the R-PE fluorescence intensity. The detection method took within 50 min with a limit of detection (LOD) was 0.05 μg/L and range from 0.2 to 20 μg/L for OA detection. Moreover, the matrix effect and the recovery rate were assessed during real sample measurement, showing a high recovery. Performance features such as high sensitivity, low LOD, speediness and simplicity of the analysis protocol, shows this biosensing-systems as a promising tool for routine use.

Keywords

Fluorescence immunosensor Portable flow cytometry Magnetic beads-based flow cytometry In-situ detection Okadaic acid 

Notes

Acknowledgements

This work was supported by a key project of the Natural Science Foundation of China (No. 31627801), International Cooperation Project of Natural Science Foundation of China (No. 61320106002,31661143030,), National 973 Project of China (No.2015CB352101), and Natural Science Foundation of China (No. 31571004).

References

  1. R. Andrew, Clinical measurement of steroid metabolism. Best Pract. Res. Clin. Endocrinol. Metab. 15, 1–16 (2001)CrossRefGoogle Scholar
  2. L.B. Bangs, New developments in particle-based immunoassays: Introduction. Pure Appl. Chem. 68, 1873–1879 (1996)CrossRefGoogle Scholar
  3. U. Bilitewski, Peer reviewed: Can affinity sensors be used to detect food contaminants? Anal. Chem. 72, 692 A–701 A (2000)CrossRefGoogle Scholar
  4. M. Campàs, J.-L. Marty, Enzyme sensor for the electrochemical detection of the marine toxin okadaic acid. Anal. Chim. Acta 605, 87–93 (2007)CrossRefGoogle Scholar
  5. M. Campàs, D. Szydlowska, M. Trojanowicz, J.-L. Marty, Towards the protein phosphatase-based biosensor for microcystin detection. Biosens. Bioelectron. 20, 1520–1530 (2005)CrossRefGoogle Scholar
  6. M. Campas, B. Prieto-Simón, J.-L. Marty, Biosensors to detect marine toxins: Assessing seafood safety. Talanta 72, 884–895 (2007)CrossRefGoogle Scholar
  7. S. Centi, S. Laschi, M. Mascini, Improvement of analytical performances of a disposable electrochemical immunosensor by using magnetic beads. Talanta 73, 394–399 (2007)CrossRefGoogle Scholar
  8. EFSA, Opinion of the scientific panel on contaminants in the food chain on a request from the European Commission on marine biotoxins in shellfish—Okadaic acid and analogues. EFSA J. 589, 1–62 (2008)Google Scholar
  9. J.V. Forment, S.P. Jackson, A flow cytometry-based method to simplify the analysis and quantification of protein association to chromatin in mammalian cells. Nat. Protoc. 10, 1297–1307 (2015)CrossRefGoogle Scholar
  10. F. Gessler, K. Hampe, M. Schmidt, H. Böhnel, Immunomagnetic beads assay for the detection of botulinum neurotoxin types C and D. Diagn. Microbiol. Infect. Dis. 56, 225–232 (2006)CrossRefGoogle Scholar
  11. H. Goto, T. Igarashi, M. Yamamoto, M. Yasuda, R. Sekiguchi, M. Watai, et al., Quantitative determination of marine toxins associated with diarrhetic shellfish poisoning by liquid chromatography coupled with mass spectrometry. J. Chromatogr. A 907, 181–189 (2001)CrossRefGoogle Scholar
  12. J.L. Guesdon, T. Ternynck, S. Avrameas, J.L. Guesdon, J. Ternyck, S. Avrameas, The use of avidin-biotin interaction in immunoenzymatic techniques. J. Histochem. Cytochem. 27, 1131–1139 (1979)CrossRefGoogle Scholar
  13. B.I. Haukanes, C. Kvam, Application of magnetic beads in bioassays. Bio/Technology 11, 60–63 (1993)Google Scholar
  14. A. Hayat, L. Barthelmebs, J.-L. Marty, Enzyme-linked immunosensor based on super paramagnetic nanobeads for easy and rapid detection of okadaic acid. Anal. Chim. Acta 690, 248–252 (2011)CrossRefGoogle Scholar
  15. A. Hayat, L. Barthelmebs, A. Sassolas, J.-L. Marty, Development of a novel label-free amperometric immunosensor for the detection of okadaic acid. Anal. Chim. Acta 724, 92–97 (2012)CrossRefGoogle Scholar
  16. T. Jung, U. Schauer, C. Heusser, C. Neumann, C. Rieger, Detection of intracellular cytokines by flow cytometry. J. Immunol. Methods 159, 197–207 (1993)CrossRefGoogle Scholar
  17. M.P. Kreuzer, C.K. O'Sulliva, G.G. Guilbault, Development of an ultrasensitive immunoassay for rapid measurement of okadaic acid and its isomers. Anal. Chem. 71, 4198–4202 (1999)CrossRefGoogle Scholar
  18. N.M. Llamas, L. Stewart, T. Fodey, H.C. Higgins, M.L.R. Velasco, L.M. Botana, et al., Development of a novel immunobiosensor method for the rapid detection of okadaic acid contamination in shellfish extracts. Anal. Bioanal. Chem. 389, 581–587 (2007)CrossRefGoogle Scholar
  19. A.P. Louppis, A.V. Badeka, P. Katikou, et al., Determination of okadaic acid, dinophysistoxin-1 and related esters in Greek mussels using HPLC with fluorometric detection, LC-MS/MS and mouse bioassay. Toxicon 55(4), 724–733 (2010)CrossRefGoogle Scholar
  20. B. Prieto-Simón, H. Miyachi, I. Karube, H. Saiki, High-sensitive flow-based kinetic exclusion assay for okadaic acid assessment in shellfish samples. Biosens. Bioelectron. 25, 1395–1401 (2010)CrossRefGoogle Scholar
  21. C. Riccardi, I. Nicoletti, Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat. Protoc. 1, 1458–1461 (2006)CrossRefGoogle Scholar
  22. S. Siena, M. Bregni, B. Brando, N. Belli, F. Ravagnani, L. Gandola, et al., Flow cytometry for clinical estimation of circulating hematopoietic progenitors for autologous transplantation in cancer patients. Blood 77, 400–409 (1991)Google Scholar
  23. K. Su, X. Qiu, J. Fang, Q. Zou, P. Wang, An improved efficient biochemical detection method to marine toxins with a smartphone-based portable system − bionic e-eye. Sensors Actuators B Chem. 238, 1165–1172 (2016)CrossRefGoogle Scholar
  24. A.X. Tang, M. Pravda, G.G. Guilbault, S. Piletsky, A.P. Turner, Immunosensor for okadaic acid using quartz crystal microbalance. Anal. Chim. Acta 471, 33–40 (2002)CrossRefGoogle Scholar
  25. T. Yasumoto, M. Murata, Marine toxins. Chem. Rev. 93, 1897–1909 (1993)CrossRefGoogle Scholar
  26. L. Zou, Q. Wang, M. Tong, H. Li, J. Wang, N. Hu, et al., Detection of diarrhetic shellfish poisoning toxins using high-sensitivity human cancer cell-based impedance biosensor. Sensors Actuators B Chem. 222(, 205–212 (2016)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biosensor National Special Laboratory, Key Laboratory for Biomedical Engineering of Education Ministry, Department of Biomedical EngineeringZhejiang UniversityHangzhouChina
  2. 2.State Key Laboratory of Transducer TechnologyChinese Academy of SciencesShanghaiChina
  3. 3.Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Optoelectronic EngineeringShenzhen UniversityShenzhenChina
  4. 4.Department of Medicine, Biomaterials Innovation Research Center, Center for Biomedical EngineeringBrigham and Women’s Hospital, Harvard Medical SchoolCambridgeUSA

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