Optimized magnetic bead-based immunoassay for automated detection of protein toxins

Abstract

Rapid, accurate, and autonomous analysis of bioagents in the environment is critical in protecting human health from natural and intentional environmental contamination with biological toxicants. We previously developed and tested an immunoassay protocol that can be utilized for automated and simultaneous detection of selected biological agents and toxins. We adopted an antibody-based approach for the detection of pathogens and/or toxins. The fluorescent eTags™ were used as reporter molecules and the immunoassay was modified for automated field-deployed detection of pathogens and/or toxins. The present study improved the limit of detection of this system to be suitable for the detection of environmental toxins. We tested different settings to optimize the assay protocol and successfully detected 10 ng/mL (or 100 fg) of an toxin analog, ovalbumin. The developed assay represents a notable improvement from currently available assays in terms of reduced time, increased sensitivity, and automation potential. Additionally, this assay can be easily modified, with the appropriate antibodies, to detect a wide range of proteins and infectious agents.

References

  1. 1.

    An, Y.R. et al. Analysis of toxicity of tetrabutyltin: comparing with EDC chemicals. Mol. Cell. Toxicol. 7, 95–101 (2011).

    Article  CAS  Google Scholar 

  2. 2.

    Yim, W.C. et al. Identification of novel 17β-estradiol (E2) target genes using cross-experiment gene expression datasets. Toxicol. Environ. Health Sci. 2, 25–38 (2010).

    Article  Google Scholar 

  3. 3.

    Ko, Y.J. et al. Field application of a recombinant protein-based ELISA during the 2010 outbreak of footand-mouth disease type A in South Korea. J. Virol. Methods 179, 265–268 (2012).

    Article  CAS  Google Scholar 

  4. 4.

    Park, T.J. et al. Development of label-free optical diagnosis for sensitive detection of influenza virus with genetically engineered fusion protein. Talanta 30, 246–252 (2012).

    Article  Google Scholar 

  5. 5.

    Kwon, Y. et al. Magnetic bead based immunoassay for autonomous detection of toxins. Anal. Chem. 80, 8416–8423 (2008).

    Article  CAS  Google Scholar 

  6. 6.

    Kang, J., Kim, S. & Kwon, Y. Antibody based biosensors for environmental monitoring. Toxicol. Environ. Health Sci. 1, 145–150 (2009).

    Article  Google Scholar 

  7. 7.

    McBride, M.T. et al. Autonomous detection of aerosolized Bacillus anthracis and Yersinia pestis. Anal. Chem. 75, 5293–5299 (2003).

    Article  CAS  Google Scholar 

  8. 8.

    Hindson, B.J. et al. Development of an automated sample preparation module for environmental monitoring of biowarfare agents. Anal. Chem. 76, 3492–3497 (2004).

    Article  CAS  Google Scholar 

  9. 9.

    Hindson, B.J. et al. Autonomous detection of aerosolized biological agents by multiplexed immunoassay with polymerase chain reaction confirmation. Anal. Chem. 77, 284–289 (2005).

    Article  CAS  Google Scholar 

  10. 10.

    Yim, W.C., Min, K., Jung, D., Lee, B.M. & Kwon, Y. Cross experimental analysis of microarray gene expression data from volatile organic compounds treated targets. Mol. Cell. Toxicol. 7, 233–241 (2011).

    Article  CAS  Google Scholar 

  11. 11.

    Chan-Hui, P.Y., Stephens, K., Warnock, R.A. & Singh, S. Applications of eTag trade mark assay platform to systems biology approaches in molecular oncology and toxicology studies. Clin. Immunol. 111, 162–174 (2004).

    Article  CAS  Google Scholar 

  12. 12.

    Wen, J., Geng, Z., Yin, Y. & Wang, Z. Versatile water soluble fluorescent probe for ratiometric sensing of Hg2+ and bovine serum albumin. Dalton Trans 14, 9737–9745 (2011).

    Article  Google Scholar 

  13. 13.

    Zeni, O. & Scarf, M.R. DNA damage by carbon nanotubes using the single cell gel electrophoresis technique. Methods Mol. Biol. 625, 109–119 (2010).

    Article  CAS  Google Scholar 

  14. 14.

    Walczak, R. et al. Fluorescence detection by miniaturized instrumentation based on non-cooled CCD minicamera and dedicated for lab-on-a-chip applications. BioChip J. 5, 271–279 (2011).

    Article  CAS  Google Scholar 

  15. 15.

    Kwon, Y., Han, Z., Karatan, E., Mrksich, M. & Kay, B. Antibody arrays prepared by cutinase-mediated immobilization on self-assembled monolayers. Anal. Chem. 76, 5713–5720 (2004).

    Article  CAS  Google Scholar 

  16. 16.

    Lionnet, T. et al. Magnetic trap construction. Cold Spring Harb. Protoc. 1, 133–138 (2012).

    Google Scholar 

  17. 17.

    Pai, N.P. & Pai, M. Point-of-care diagnostics for HIV and tuberculosis: landscape, pipeline, and unmet needs. Discov. Med. 13, 35–45 (2012).

    Google Scholar 

  18. 18.

    Friedman, E.S. et al. A cost-effective and field-ready potentiostat that poises subsurface electrodes to monitor bacterial respiration. Biosens. Bioelectron. 32, 309–313 (2012).

    Article  CAS  Google Scholar 

  19. 19.

    Inglis, T.J. et al. Deployable laboratory response to influenza pandemic; PCR assay field trials and comparison with reference methods. PLoS One. 6, e25526 (2011).

    Article  CAS  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Youngeun Kwon.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jung, DH., Min, K., Jeon, Y. et al. Optimized magnetic bead-based immunoassay for automated detection of protein toxins. BioChip J 6, 293–298 (2012). https://doi.org/10.1007/s13206-012-6312-3

Download citation

Keywords

  • Immunoassay
  • Magnetic bead
  • Autonomous detection
  • Toxins
  • Fluorescent tag