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Solid-phase synthesis of molecularly imprinted nanoparticles

Nature Protocols volume 11, pages 443–455 (2016)Cite this article

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Abstract

Molecularly imprinted polymers (MIPs) are synthetic materials, generally based on acrylic or methacrylic monomers, that are polymerized in the presence of a specific target molecule called the 'template' and capable of rebinding selectively to this target molecule. They have the potential to be low-cost and robust alternatives to biomolecules such as antibodies and receptors. When prepared by traditional synthetic methods (i.e., with free template in solution), their usefulness has been limited by high binding site heterogeneity, the presence of residual template and the fact that the production methods are complex and difficult to standardize. To overcome some of these limitations, we developed a method for the synthesis of MIP nanoparticles (nanoMIPs) using an innovative solid-phase approach, which relies on the covalent immobilization of the template molecules onto the surface of a solid support (glass beads). The obtained nanoMIPs are virtually free of template and demonstrate high affinity for the target molecule (e.g., melamine and trypsin in our published work). Because of an affinity separation step performed on the solid phase after polymerization, poor binders and unproductive polymer are removed, so the final product has more uniform binding characteristics. The overall protocol, starting from the immobilization of the template onto the solid phase and including the purification and characterization of the nanoparticles, takes up to 1 week.

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Figure 1: Immobilization of template molecules on the solid support, for three common functional groups.
Figure 2: Solid-phase synthesis and separation of nanoMIPs by photo-polymerization, using melamine as a model template (for illustration purposes).
Figure 3: Overview of the process for the production of nanoMIPs by solid-phase imprinting.

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Acknowledgements

This research was supported by the Wellcome Trust with a Translation Award and by the Research Executive Agency (REA) of the European Union under grant agreement number PITN-GA-2010-264772 (ITN CHEBANA). A.P. is a recipient of the National Centre for the Replacement, Refinement & Reduction of Animals in Research (NC3Rs) David Sainsbury Fellowship.

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Authors and Affiliations

  1. MIP Diagnostics, University of Leicester, Leicester, UK

    Francesco Canfarotta

  2. Department of Chemistry, University College London, London, UK

    Alessandro Poma

  3. Department of Chemistry, University of Leicester, Leicester, UK

    Antonio Guerreiro & Sergey Piletsky

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  1. Francesco Canfarotta
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  2. Alessandro Poma
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  3. Antonio Guerreiro
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  4. Sergey Piletsky
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Contributions

F.C., A.P., A.G. and S.P. developed the protocols. F.C., A.G. and A.P. contributed the data and wrote the paper. All authors have discussed the results and approved the final version of the manuscript.

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Correspondence to Francesco Canfarotta.

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The authors declare no competing financial interests.

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Canfarotta, F., Poma, A., Guerreiro, A. et al. Solid-phase synthesis of molecularly imprinted nanoparticles. Nat Protoc 11, 443–455 (2016). https://doi.org/10.1038/nprot.2016.030

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