Abstract
Synthetic polymer nanoparticles (NPs) with intrinsic affinity for target biomacromolecules hold great promise in the development of novel tools for biological and biomedical research. We recently reported the design and synthesis of abiotic, synthetic polymer NPs with high intrinsic affinity for a peptide toxin melittin. The NP was selected by screening a small library of NPs (ā¼100 nm) composed of various ratios of monomers that contain functional groups complementary to the peptide melittin. The selected polymer NP, a co-polymer of acrylic acid (AAc), N-tert-butylacrylamide (TBAm), N-isopropylacrylamide (NIPAm) and N,Nā²-methylenebisacrylamide (BIS), effectively captures and neutralizes the toxicity of the peptide through a combination of electrostatic and hydrophobic interactions. This protocol describes a step-by-step procedure for the preparation and evaluation of synthetic polymer NPs for sequestration and neutralization of the target peptide toxin. The polymer NPs can be synthesized in a one-step polymerization reaction using commercially available reagents. The polymerization reaction for the synthesis of polymer NPs takes several hours, and the total protocol including subsequent purification and characterization by dynamic light scattering, NMR and toxicity neutralization assays takes 1ā2 weeks in total.
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Acknowledgements
This research was supported by the US National Science Foundation (DMR-1308363). H.K. is a recipient of the Japan Society for the Promotion of Science (JSPS) Fellowship.
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K.Y., H.K., Y.H. and K.J.S. developed protocols. K.Y. and H.K. contributed all the data. K.Y., H.K. and K.J.S. wrote the paper. All authors have discussed the results and approved the final manuscript.
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Supplementary Text and Figures
Supplementary Table 1 (PDF 88 kb)
Supplementary Data 1
1H NMR spectrum (500 MHz, CD3OD) of polymer NP synthesized by the copolymerization of 5 mol% AAc, 40 mol% TBAm, 53 mol% NIPAm, and 2 mol% BIS (values are in monomer feed ratio). (ZIP 88 kb)
Supplementary Data 2
13C NMR spectrum (126 MHz, CD3OD) of polymer NP synthesized by the copolymerization of 5 mol% AAc, 40 mol% TBAm, 53 mol% NIPAm, and 2 mol% BIS (values are in monomer feed ratio). (ZIP 225 kb)
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Yoshimatsu, K., Koide, H., Hoshino, Y. et al. Preparation of abiotic polymer nanoparticles for sequestration and neutralization of a target peptide toxin. Nat Protoc 10, 595ā604 (2015). https://doi.org/10.1038/nprot.2015.032
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DOI: https://doi.org/10.1038/nprot.2015.032
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