Analytical and Bioanalytical Chemistry

, Volume 408, Issue 18, pp 5013–5019 | Cite as

Magnetic-bead-based sub-femtomolar immunoassay using resonant Raman scattering signals of ZnS nanoparticles

  • Yadan Ding
  • Tie Cong
  • Xueying Chu
  • Yan Jia
  • Xia HongEmail author
  • Yichun LiuEmail author
Research Paper


Highly sensitive, specific, and selective immunoassays are of great significance for not only clinical diagnostics but also food safety, environmental monitoring, and so on. Enzyme-linked immunosorbent assays and fluorescence-based and electrochemical immunoassays are important intensively investigated immunoassay techniques. However, they might suffer from low sensitivity or false-positive results. In this work, a simple, reliable, and ultrasensitive magnetic-bead-based immunoassay was performed using biofunctionalized ZnS semiconductor nanocrystals as resonant Raman probes. The resonant Raman scattering of ZnS nanocrystals displays evenly spaced multi-phonon resonant Raman lines with narrow bandwidths and has strong resistance to environmental variation due to the nature of the electron-phonon interaction, thus rendering reliable signal readout in the immunoassays. The superparamagnetic Fe3O4 nanoparticles facilitated greatly the separation, purification, and concentration processes. It is beneficial for both reducing the labor intensity and amplifying the detection signals. The immobilization of antibodies on the surface of magnetic beads, the preparation of resonant Raman probes, and the immunological recognition between the antibody and analyte all occurred in the liquid phase, which minimized the diffusion barriers and boundary layer constraints. All these factors contributed to the ultralow detection limit of human IgG, which was determined to be about 0.5 fM (∼0.08 pg/ml). It is nearly the highest sensitivity obtained for IgG detection. This work shall facilitate the design of nanoplatforms for ultrasensitive detections of proteins, DNAs, bacteria, explosives, and so on.

Graphical abstract

An ultrasensitive magnetic-bead-based immunoassay was performed using multi-phonon resonant Raman lines of ZnS nanoparticles as detection signals


Magnetic bead ZnS nanocrystal Resonant Raman scattering Immunoassay 



This work was supported by the National Natural Science Foundation of China (grants no. 51272040, 51072031, 11304035, and 61205193), the 111 project (no. B13013), and the Program for New Century Excellent Talents in the University of Ministry of Education of China (grant no. NCET-12-0815).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    Le T, Yan P, Xu J, Hao Y. A novel colloidal gold-based lateral flow immunoassay for rapid simultaneous detection of cyromazine and melamine in foods of animal origin. Food Chem. 2013;138(2–3):1610–5.CrossRefGoogle Scholar
  2. 2.
    Giljohann DA, Mirkin CA. Drivers of biodiagnostic development. Nature. 2009;462(7272):461–4.CrossRefGoogle Scholar
  3. 3.
    Zhan L, Li CM, Wu WB, Huang CZ. A colorimetric immunoassay for respiratory syncytial virus detection based on gold nanoparticles-graphene oxide hybrids with mercury-enhanced peroxidase-like activity. Chem Commun. 2014;50(78):11526–8.CrossRefGoogle Scholar
  4. 4.
    Shen J, Li Y, Gu H, Xia F, Zuo X. Recent development of sandwich assay based on the nanobiotechnologies for proteins, nucleic acids, small molecules, and ions. Chem Rev. 2014;114(15):7631–77.CrossRefGoogle Scholar
  5. 5.
    Pedrero M, Campuzano S, Pingarron JM. Magnetic beads-based electrochemical sensors applied to the detection and quantification of bioterrorism/biohazard agents. Electroanalysis. 2012;24(3):470–82.CrossRefGoogle Scholar
  6. 6.
    Peterson RD, Chen W, Cunningham BT, Andrade JE. Enhanced sandwich immunoassay using antibody-functionalized magnetic iron-oxide nanoparticles for extraction and detection of soluble transferrin receptor on a photonic crystal biosensor. Biosens Bioelectron. 2015;74:815–22.CrossRefGoogle Scholar
  7. 7.
    Huang X, Li Y, Huang X, Xie X, Xu Y, Chen Y, et al. A novel reverse fluorescent immunoassay approach for sensing human chorionic gonadotropin based on silver-gold nano-alloy and magnetic nanoparticles. Anal Bioanal Chem. 2016;408(2):619–27.CrossRefGoogle Scholar
  8. 8.
    Chen Y, Cheng H, Tram K, Zhang S, Zhao Y, Han L, et al. A paper-based surface-enhanced resonance Raman spectroscopic (SERRS) immunoassay using magnetic separation and enzyme-catalyzed reaction. Analyst. 2013;138(9):2624–31.CrossRefGoogle Scholar
  9. 9.
    Zhang J, Wang S, Liu K, Wei Y, Wang X, Duan Y. Novel signal-enhancing immunoassay for ultrasensitive biomarker detection based on laser-induced fluorescence. Anal Chem. 2015;87(5):2959–65.CrossRefGoogle Scholar
  10. 10.
    Li C-L, Huang B-R, Chang J-Y, Chen J-K. Bifunctional superparamagnetic-luminescent core-shell-satellite structured microspheres: preparation, characterization, and magnetodisplay application. J Mater Chem C. 2015;3(18):4603–15.CrossRefGoogle Scholar
  11. 11.
    Hong X, Li J, Wang M, Xu J, Guo W, Li J, et al. Fabrication of magnetic luminescent nanocomposites by a layer-by-layer self-assembly approach. Chem Mater. 2004;16:4022–7.CrossRefGoogle Scholar
  12. 12.
    Stanisavljevic M, Krizkova S, Vaculovicova M, Kizek R, Adam V. Quantum dots-fluorescence resonance energy transfer-based nanosensors and their application. Biosens Bioelectron. 2015;74:562–74.CrossRefGoogle Scholar
  13. 13.
    Zhou J, Yang Y, Zhang C-y. Toward biocompatible semiconductor quantum dots: from biosynthesis and bioconjugation to biomedical application. Chem Rev. 2015;115(21):11669–717.CrossRefGoogle Scholar
  14. 14.
    Milekhin AG, Yeryukov NA, Sveshnikova LL, Duda TA, Himcinschi C, Zenkevich EI, et al. Resonant Raman scattering of ZnS, ZnO, and ZnS/ZnO core/shell quantum dots. Appl Phys A-Mater. 2012;107(2):275–8.CrossRefGoogle Scholar
  15. 15.
    Sahoo S, Arora AK. Laser-power-induced multiphonon resonant Raman scattering in laser-heated CdS nanocrystal. J Phys Chem B. 2010;114(12):4199–203.CrossRefGoogle Scholar
  16. 16.
    Hong X, Chu X, Zou P, Liu Y, Yang G. Magnetic-field-assisted rapid ultrasensitive immunoassays using Fe3O4/ZnO/Au nanorices as Raman probes. Biosens Bioelectron. 2010;26(2):918–22.CrossRefGoogle Scholar
  17. 17.
    Sun S, Zeng H, Robinson DB, Raoux S, Rice PM, Wang SX, et al. Monodisperse MFe2O4 (M = Fe, Co, Mn) nanoparticles. J Am Chem Soc. 2004;126(1):273–9.CrossRefGoogle Scholar
  18. 18.
    Duan H, Kuang M, Wang X, Wang YA, Mao H, Nie S. Reexamining the effects of particle size and surface chemistry on the magnetic properties of iron oxide nanocrystals: new insights into spin disorder and proton relaxivity. J Phys Chem C. 2008;112(22):8127–31.CrossRefGoogle Scholar
  19. 19.
    Chu X, Hong X, Zou P, Men J, Liu Y. Ultrasensitive protein detection in terms of multiphonon resonance Raman scattering in ZnS nanocrystals. Appl Phys Lett. 2011;98(25):253073.Google Scholar
  20. 20.
    Zhu Y, Zhao W, Chen H, Shi J. A simple one-pot self-assembly route to nanoporous and monodispersed Fe3O4 particles with oriented attachment structure and magnetic property. J Phys Chem C. 2007;111(14):5281–5.CrossRefGoogle Scholar
  21. 21.
    Mandal SK, Mandal AR, Das S, Bhattacharjee B. Strong excitonic confinement effect in ZnS and ZnS: Mn nanorods embedded in polycarbonate membrane pores. J Appl Phys. 2007;101(11):114315.CrossRefGoogle Scholar
  22. 22.
    Davar F, Mohammadikish M, Loghman-Estarki MR, Hamidi Z. Synthesis of spherical ZnS based nanocrystals using thioglycolic assisted hydrothermal method. CrystEngComm. 2012;14(21):7338–44.CrossRefGoogle Scholar
  23. 23.
    Luo YY, Duan GT, Li GH. Resonant Raman scattering and surface phonon modes of hollow ZnS microspheres. Appl Phys Lett. 2007;90(20):201911.CrossRefGoogle Scholar
  24. 24.
    Brafman O, Mitra SS. Raman effect in wurtzite- and zinc-blende-type ZnS single crystals. Phys Rev. 1968;171:931.CrossRefGoogle Scholar
  25. 25.
    Wang S, Chen Z, Choo J, Chen L. Naked-eye sensitive ELISA-like assay based on gold-enhanced peroxidase-like immunogold activity. Anal Bioanal Chem. 2016;408(4):1015–22.CrossRefGoogle Scholar
  26. 26.
    Kumari S, Dhar BB, Panda C, Meena A, Sen GS. Fe-TAML encapsulated inside mesoporous silica nanoparticles as peroxidase mimic: femtomolar protein detection. ACS Appl Mater Interfaces. 2014;6(16):13866–73.CrossRefGoogle Scholar
  27. 27.
    Zhang S, Huang N, Lu Q, Liu M, Li H, Zhang Y, et al. A double signal electrochemical human immunoglobulin G immunosensor based on gold nanoparticles-polydopamine functionalized reduced graphene oxide as a sensor platform and AgNPs/carbon nanocomposite as signal probe and catalytic substrate. Biosens Bioelectron. 2016;77:1078–85.CrossRefGoogle Scholar
  28. 28.
    Li R, Wu K, Liu C, Huang Y, Wang Y, Fang H, et al. 4-Amino-1-(3-mercapto-propyl)-pyridine hexafluorophosphate ionic liquid functionalized gold nanoparticles for IgG immunosensing enhancement. Anal Chem. 2014;86(11):5300–7.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Key Laboratory of UV-Emitting Materials and Technology (Northeast Normal University)Ministry of EducationChangchunChina
  2. 2.School of ScienceChangchun University of Science and TechnologyChangchunChina

Personalised recommendations