An Ultrasensitive Biosensing Platform Employing Acetylcholinesterase and Gold Nanoparticles

  • Dingbin LiuEmail author
  • Xiaoyuan Chen
Part of the Methods in Molecular Biology book series (MIMB, volume 1530)


Enzyme-linked immunosorbent assay (ELISA) is a well-known strategy for biomarker detection with a color change, which can be seen by the naked eyes. However, the moderate sensitivity of conventional ELISA limits its applications in many cases where the concentrations of biomarker are very low, such as cancer diagnosis. Here we describe an ultrasensitive colorimetric assay based on acetylcholinesterase (AChE)-catalyzed hydrolysis reaction, whose products trigger the aggregation of gold nanoparticles (AuNPs), causing a distinct color change of the solution from red to purple. This enhanced colorimetric immunoassay offers extremely high sensitivity and specificity. In this study, we employed enterovirus 71 (EV71), the major cause of hand, foot, and mouth disease (HFMD), as a model to evaluate the analytical performance of the plasmonic immunoassay.

Key words

Gold nanoparticles Colorimetric immunoassay Acetylcholinesterase Magnetic beads Signal amplification 



We acknowledge the support from the National Natural Science Foundation of China (Grants 21475066 and 81401463), the Fundamental Research Funds for Central Universities (China), and the Intramural Research Program (IRP) of the National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health.


  1. 1.
    Park JS, Cho MK, Lee EJ, Ahn KY, Lee KE, Jung JH et al (2009) A highly sensitive and selective diagnostic assay based on virus nanoparticles. Nat Nanotechnol 4:259–264CrossRefPubMedGoogle Scholar
  2. 2.
    Rissin DM, Kan CW, Campbell TG, Howes SC, Fournier DR, Song L et al (2010) Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations. Nat Biotechnol 28:595–599CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Rosi NL, Mirkin CA (2005) Nanostructures in biodiagnostics. Chem Rev 105:1547–1562CrossRefPubMedGoogle Scholar
  4. 4.
    Liu D, Qu W, Chen W, Zhang W, Wang Z, Jiang X (2010) Highly sensitive, colorimetric detection of mercury(II) in aqueous media by quaternary ammonium group-capped gold nanoparticles at room temperature. Anal Chem 82:9606–9610CrossRefPubMedGoogle Scholar
  5. 5.
    Zagorovsky K, Chan WCW (2013) A plasmonic DNAzyme strategy for point-of-care genetic detection of infectious pathogens. Angew Chem Int Ed 52:3168–3171CrossRefGoogle Scholar
  6. 6.
    Liu JW, Lu Y (2006) Fast colorimetric sensing of adenosine and cocaine based on a general sensor design involving aptamers and nanoparticles. Angew Chem Int Ed 45:90–94CrossRefGoogle Scholar
  7. 7.
    De la Rica R, Stevens MM (2012) Plasmonic ELISA for the ultrasensitive detection of disease biomarkers with the naked eye. Nat Nanotechnol 7:821–824CrossRefPubMedGoogle Scholar
  8. 8.
    Liu D, Wang Z, Jin A, Huang X, Sun X, Wang F et al (2013) Acetylcholinesterase-catalyzed hydrolysis allows ultrasensitive detection of pathogens with the naked eye. Angew Chem Int Ed 52:14065–14069CrossRefGoogle Scholar
  9. 9.
    Nie XM, Huang R, Dong CX, Tang LJ, Gui R, Jiang JH (2014) Plasmonic ELISA for the ultrasensitive detection of treponema pallidum. Biosens Bioelectron 58:314–319CrossRefPubMedGoogle Scholar
  10. 10.
    Marks IS, Kang JS, Jones BT, Landmark KJ, Cleland AJ, Taton TA (2011) Strain-promoted “click” chemistry for terminal labeling of DNA. Bioconjug Chem 22:1259–1263CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Wang M, Gu XG, Zhang GX, Zhang DQ, Zhu DB (2009) Continuous colorimetric assay for acetylcholinesterase and inhibitor screening with gold nanoparticles. Langmuir 25:2504–2507CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.College of Chemistry, Research Center for Analytical Sciences, State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Molecular Recognition and Biosensing, and Collaborative Innovation Center of Chemical Science and EngineeringNankai UniversityTianjinChina
  2. 2.Laboratory of Molecular Imaging and Nanomedicine (LOMIN)National Institute of Biomedical Imaging and Bioengineering (NIBIB), National Institutes of Health (NIH)BethesdaUSA

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