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A molecular-imprint nanosensor for ultrasensitive detection of proteins

Nature Nanotechnology volume 5, pages 597–601 (2010)Cite this article

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Abstract

Molecular imprinting is a technique for preparing polymer scaffolds that function as synthetic receptors1,2,3. Imprinted polymers that can selectively bind organic compounds have proven useful in sensor development2,3,4,5,6,7. Although creating synthetic molecular-imprinting polymers that recognize proteins remains challenging8,9,10,11, nanodevices and nanomaterials show promise in this area12,13,14. Here, we show that arrays of carbon-nanotube tips with an imprinted non-conducting polymer coating can recognize proteins with subpicogram per litre sensitivity using electrochemical impedance spectroscopy. We have developed molecular-imprinting sensors specific for human ferritin and human papillomavirus derived E7 protein. The molecular-imprinting-based nanosensor can also discriminate between Ca2+-induced conformational changes in calmodulin. This ultrasensitive, label-free electrochemical detection of proteins offers an alternative to biosensors based on biomolecule recognition.

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Figure 1: Fabrication of an MI protein nanosensor.
Figure 2: Detection of hFtn using an MI nanosensor.
Figure 3: Detection mechanism of the MI nanosensor.
Figure 4: Detection of calcium-dependent calmodulin conformational changes and HPV-derived oncoproteins.

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References

  1. Wulff, G. & Sharhan, A. Über die Anwendung von enzymanalog gebauten Polymeren zur Racemattrennung. Angew. Chem. 84, 364 (1972).

    Article  Google Scholar 

  2. Vlatakis, G., Anderson, L. I., Muller, R. & Mosbach, K. Drug assay using antibody mimics made by molecular imprinting. Nature 361, 645–647 (1993).

    Article  CAS  Google Scholar 

  3. Hoshino, Y., Kodama, T., Okahata, Y. & Shea, K. J. Peptide imprinted polymer nanoparticles: a plastic antibody. J. Am. Chem. Soc. 130, 15242–15243 (2008).

    Article  CAS  Google Scholar 

  4. Bossi, A., Bonini, F., Turner, A. P. F. & Piletsky, S. A. Molecularly imprinted polymers for the recognition of proteins: the state of the art. Biosens. Bioelectron. 22, 1131–1137 (2007).

    Article  CAS  Google Scholar 

  5. Karube, J. et al. Molecular recognition in continuous polymer rods prepared by a molecular imprinting technique. Anal. Chem. 65, 2223–2224 (1993).

    Article  Google Scholar 

  6. Kempe, M. Antibody-mimicking polymers as chiral stationary phases in HPLC. Anal. Chem. 68, 1948–1953 (1996).

    Article  CAS  Google Scholar 

  7. Rashid, B. A., Briggs, R. J., Hay, J. N. & Stevenson, D. Preliminary evaluation of a molecular imprinted polymer for solid-phase extraction of tamoxifen. Anal. Commun. 34, 303–305 (1997).

    Article  CAS  Google Scholar 

  8. Shi, H., Tsai, W.-B., Garrison, M. D., Ferrari, S. & Ratner, B. D. Template-imprinted nanostructured surfaces for protein recognition. Nature 398, 593–597 (1999).

    Article  CAS  Google Scholar 

  9. Rick, J. & Chou, T. C. Using protein templates to direct the formation of thin-film polymer surfaces. Biosens. Bioelectron. 22, 544–549 (2006).

    Article  CAS  Google Scholar 

  10. Wang, Y. et al. A potentiometric protein sensor built with surface molecular imprinting method. Biosens. Bioelectron. 24, 162–166 (2008).

    Article  Google Scholar 

  11. Bossi, A., Piletsky, S. A., Piletska, E. V., Righetti, P. G. & Turner, A. P. F. Surface-grafted molecularly imprinted polymers for protein recognition. Anal. Chem. 73, 5281–5286 (2001).

    Article  CAS  Google Scholar 

  12. Zheng, G., Patolsky, F., Cui, Y., Wang, W. U. & Lieber, C. M. Multiplexed electrical detection of cancer markers with nanowire sensor arrays. Nature Biotechnol. 23, 1294–1301 (2005).

    Article  CAS  Google Scholar 

  13. Yu, X. et al. Carbon nanotube amplification strategies for highly sensitive immunodetection of cancer biomarkers. J. Am. Chem. Soc. 128, 11199–11205 (2006).

    Article  CAS  Google Scholar 

  14. Yu, Y. et al. Assembly of multi-functional nanocomponents on periodic nanotube array for biosensors. Micro. Nano. Lett. 4, 27–33 (2009).

    Article  CAS  Google Scholar 

  15. Riskin, M., Tel-Vered, R. & Willner, I. The imprint of electropolymerized polyphenol films on electrodes by donor–acceptor interactions: selective electrochemical sensing of N,N′-dimethyl-4,4′-bipyridinium (methyl viologen). Adv. Func. Mater. 17, 3858–3863 (2007).

    Article  CAS  Google Scholar 

  16. Lakshmi, D. et al. Electrochemical sensor for catechol and dopamine based on a catalytic molecularly imprinted polymer-conducting polymer hybrid recognition element. Anal. Chem. 81, 3576–3584 (2009).

    Article  CAS  Google Scholar 

  17. Panasyuk, T. L., Mirsky, V. M., Piletsky, S. A. & Wolfbeis, O. S. Electropolymerized molecularly imprinted polymers as receptor layers in capacitive chemical sensors. Anal. Chem. 71, 4609–4613 (1999).

    Article  CAS  Google Scholar 

  18. Choong, C. L., Bendallb, J. S. & Milnea, W. I. Carbon nanotube array: a new MIP platform. Biosen. Bioelectron. 25, 652–656 (2009).

    Article  CAS  Google Scholar 

  19. Wei, Y., Qiu, L., Owen, C. & Lai, E. P. C. Encapsulation of quantum dots and carbon nanotubes with polypyrrole in a syringe needle for automated molecularly imprinted solid phase pre-concentration of ochratoxin A in red wine analysis. Sens. Instrum. Food Qual. 1, 133–141 (2007).

    Article  Google Scholar 

  20. Xie, C., Li, H., Li, S., Wu, J. & Zhang, Z. Surface molecular self-assembly for organophosphate pesticide imprinting in electropolymerized poly(p-aminothiophenol) membranes on a gold nanoparticle modified glassy carbon electrode. Anal. Chem. 82, 241–249 (2010).

    Article  CAS  Google Scholar 

  21. Tabard-Cossa, V., Trivedi, V., Wiggin, M., Jetha, N. N. & Marziali, A. Noise analysis and reduction in solid-state nanopores. Nanotechnology 18, 305505 (2007).

    Article  Google Scholar 

  22. Gong, K., Chakrabarti, S. & Dai, L. Electrochemistry at carbon nanotube electrodes: is the nanotube tip more active than the side wall? Angew. Chem. Int. Ed. 47, 5446–5450 (2008).

    Article  CAS  Google Scholar 

  23. Bartlett, P. N., Tebbutt, P. & Tyrrell, C. H. Electrochemical immobilization of enzymes. 3. Immobilization of glucose oxidase in thin films of electrochemically polymerized phenols. Anal. Chem. 64, 138–142 (1992).

    Article  CAS  Google Scholar 

  24. Klee, C. B. Conformational transition accompanying the binding of Ca2+ to the protein activator of 3′,5′-cyclic adenosine monophosphate. Biochemistry 16, 1017–1024 (1977).

    Article  CAS  Google Scholar 

  25. Crouch, T. H. & Klee, C. B. Positive cooperative binding of calcium to bovine brain calmodulin. Biochemistry 19, 3692–3698 (1980).

    Article  CAS  Google Scholar 

  26. Weinstein, H. & Mehler, E. L. Ca2+-binding and structural dynamics in the functions of calmodulins. Annu. Rev. Physiol. 56, 213–236 (1994).

    Article  CAS  Google Scholar 

  27. Devanathan, S. et al. Subpicomolar sensing of δ-opioid receptor ligands by molecular-imprinted polymers using plasmon-waveguide resonance spectroscopy. Anal. Chem. 77, 2569–2574 (2005).

    Article  CAS  Google Scholar 

  28. Patolsky, F., Zheng, G. & Lieber, C. M. Fabrication of silicon nanowire devices for ultrasensitive, label-free, real-time detection of biological and chemical species. Nature Protoc. 1, 1711–1724 (2006).

    Article  CAS  Google Scholar 

  29. Sanchez-Carbayo, M. Antibody arrays: technical considerations and clinical applications in cancer. Clin. Chem. 52, 1651–1659 (2006).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors acknowledge financial support from The Seaver Institute (T.C.C and M.J.N.). L.R. was supported by the China Scholarship Council (no. 2008677005). We thank O. Gursky and S. Jayaraman (Boston University School of Medicine, Department of Physiology and Biophysics) for use of their Aviv spectropolarimeter.

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Authors and Affiliations

  1. Department of Biology, Boston College, Chestnut Hill, 02467, Massachusetts, USA

    Dong Cai, Lu Ren, Chenjia Xu, Lu Zhang, Jeffrey H. Chuang & Thomas C. Chiles

  2. Department of Physics, Boston College, Chestnut Hill, 02467, Massachusetts, USA

    Huaizhou Zhao, Ying Yu, Hengzhi Wang, Yucheng Lan, Michael J. Naughton & Zhifeng Ren

  3. Department of Chemistry, Boston College, Chestnut Hill, 02467, Massachusetts, USA

    Mary F. Roberts

  4. Institute of Nanoscience and Nanotechnology, Central China Normal University, Wuhan, 430079, China

    Lu Ren & Ying Yu

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  1. Dong Cai
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Contributions

D.C. contributed to the original MI nanosensor concept, overall experimental design, data analysis, manuscript preparation and directing of the measurements. L.R. was responsible for the EIS recordings and contributed to sensor fabrication. H.Z. fabricated the nanotube arrays. C.X. was responsible for protein preparation and purification. Y.Y. contributed to development of the PPn nanocoating. H.W. and Y.L. helped with the high-resolution TEM image. M.F.R. was responsible for the circular dichroism measurements. L.Z. and J.H.C. were responsible for computational analysis of the interaction between the E7 protein and PPn. M.J.N. provided technical support for nanotube fabrication and assisted in manuscript editing. Z.R. contributed expertise regarding nanotube fabrication, experimental design for TEM evaluation of hFtn entrapment, and editing of the manuscript. T.C.C. contributed to the design of experiments for demonstrating nanosensor selectivity and was responsible for writing and editing the revised manuscript.

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Correspondence to Dong Cai.

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Cai, D., Ren, L., Zhao, H. et al. A molecular-imprint nanosensor for ultrasensitive detection of proteins. Nature Nanotech 5, 597–601 (2010). https://doi.org/10.1038/nnano.2010.114

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