Detection of ricin toxin in water using immunoassays

Two types of immunoassays were employed to determine ricin toxin through the substitute of A chain sub-unit in water. Protocol for detecting ricin by commercial enzyme-linked immunosorbent assay (ELISA) kits was established and evaluated. Final results were obtained by this assay in six hours and the detection limit was found to be less than 3 ng mL−1 based on ricin A chain. Cross-reactivity from ricin B chain was minimal. The validated assay was successfully applied to the determination of ricin toxin in water for the purpose of water security. The results showed that water samples were free from this biotoxin. In order to reduce the assay time and enhance the ability of securing water quality, a fibre optic biosensor RAPTOR™ has been applied as another platform to rapidly analyse ricin toxin in water. Polyclonal anti-ricin antibody was captured on the waveguide through incubation. Four waveguides were assembled into each coupon, which could provide multiplex analysis in one assay run. Cy5-labelled antibody was used as detector antibody and characterised by MALDI-TOF mass spectrometry. Results showed that the detection limit was 10 and 60 ng mL−1 of ricin A chain in deionised water and tap water, respectively. Based on readily assembled coupon, the assay run could be completed and the results reported in less than 15 minutes. Thus, the biosensor has been shown to be a promising technology for rapid and sensitive detection of biotoxins in water. In the future, more research work will be carried out to further improve the sensitivity and enhance reproducibility of the assay for the detection of ricin toxin.

[1]  U. Narang,et al.  Fiber optic-based biosensor for ricin. , 1997, Biosensors & bioelectronics.

[2]  M. Poli,et al.  Detection of ricin by colorimetric and chemiluminescence ELISA. , 1994, Toxicon : official journal of the International Society on Toxinology.

[3]  F. Ligler,et al.  Detection of multiple toxic agents using a planar array immunosensor. , 1998, Biosensors & bioelectronics.

[4]  Giyoung Kim,et al.  Antibody Immobilization on Waveguides Using a Flow–Through System Shows Improved Listeria monocytogenes Detection in an Automated Fiber Optic Biosensor: RAPTOR™ , 2006, Sensors (Basel, Switzerland).

[5]  Douglas L. Park,et al.  Feasibility of immunodiagnostic devices for the detection of ricin, amanitin, and T-2 toxin in food. , 2005, Journal of food protection.

[6]  S. Ramakrishnan,et al.  Radioimmunoassay of ricin A- and B-chains applied to samples of ricin A-chain prepared by chromatofocusing and by DEAE Bio-Gel A chromatography. , 1982, Biochimica et biophysica acta.

[7]  Frances S. Ligler,et al.  Array biosensor for detection of toxins , 2003, Analytical and bioanalytical chemistry.

[8]  George P. Anderson,et al.  Water quality monitoring using an automated portable fiber optic biosensor: RAPTOR , 2001, SPIE Optics East.

[9]  Lokendra Singh,et al.  Biological Warfare Agents , 2006 .

[10]  George P. Anderson,et al.  RAPTOR: a fluoroimmunoassay-based fiber optic sensor for detection of biological threats , 2003 .

[11]  Frances S Ligler,et al.  A microarray immunoassay for simultaneous detection of proteins and bacteria. , 2002, Analytical chemistry.

[12]  Renato Zenobi,et al.  Immunoassays with direct mass spectrometric detection. , 2006, Analytical chemistry.

[13]  D. Lim,et al.  A rapid and automated fiber optic-based biosensor assay for the detection of Salmonella in spent irrigation water used in the sprouting of sprout seeds. , 2004, Journal of food protection.

[14]  Huijie Huang,et al.  Direct detection of Yersinia pestis from the infected animal specimens by a fiber optic biosensor , 2007 .

[15]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[16]  George P. Anderson,et al.  Automated Fiber Optic Biosensor for Multiplexed Immunoassays , 2000 .

[17]  A Pihl,et al.  Radioimmunoassays of abrin and ricin in blood. , 1981, Journal of toxicology and environmental health.

[18]  N. Koja,et al.  Enzyme-linked immunoassay of ricin. , 1980, Toxicon : official journal of the International Society on Toxinology.

[19]  G. Anderson,et al.  Multi-analyte interrogation using the fiber optic biosensor. , 2000, Biosensors & bioelectronics.

[20]  Arun K. Bhunia,et al.  Fiber-Optic Biosensor Employing Alexa-Fluor Conjugated Antibody for Detection of Escherichia coli O157:H7 from Ground Beef in Four Hours , 2006, Sensors (Basel, Switzerland).

[21]  G. Shi,et al.  A water-soluble cationic oligopyrene derivative : Spectroscopic studies and sensing applications , 2009 .

[22]  M. S. Lewis,et al.  Ricin subunit association. Thermodynamics and the role of the disulfide bond in toxicity. , 1986, The Journal of biological chemistry.

[23]  F. Ligler,et al.  Evanescent wave fluorescence biosensors. , 2005, Biosensors & bioelectronics.

[24]  Hwan-wun Liu,et al.  Monoclonal antibody-based enzyme immunoassay for detection of ricin. , 2002, Hybridoma and hybridomics.

[25]  Nandan L Nerurkar,et al.  Improved fluoroimmunoassays using the dye Alexa Fluor 647 with the RAPTOR, a fiber optic biosensor. , 2002, Journal of immunological methods.

[26]  M. Thevis,et al.  Mass spectrometric characterization of transferrins and their fragments derived by reduction of disulfide bonds , 2003, Journal of the American Society for Mass Spectrometry.

[27]  J. Osterloh,et al.  Ricin poisoning: a comprehensive review. , 2005, JAMA.