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Gradated assembly of multiple proteins into supramolecular nanomaterials


Biomaterials exhibiting precise ratios of different bioactive protein components are critical for applications ranging from vaccines to regenerative medicine, but their design is often hindered by limited choices and cross-reactivity of protein conjugation chemistries. Here, we describe a strategy for inducing multiple different expressed proteins of choice to assemble into nanofibres and gels with exceptional compositional control. The strategy employs ‘βTail’ tags, which allow for good protein expression in bacteriological cultures, yet can be induced to co-assemble into nanomaterials when mixed with additional β-sheet fibrillizing peptides. Multiple different βTail fusion proteins could be inserted into peptide nanofibres alone or in combination at predictable, smoothly gradated concentrations, providing a simple yet versatile route to install precise combinations of proteins into nanomaterials. The technology is illustrated by achieving precisely targeted hues using mixtures of fluorescent proteins, by creating nanofibres bearing enzymatic activity, and by adjusting antigenic dominance in vaccines.

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Figure 1: Engineered fusion proteins with a β-sheet fibrillizing tail integrate into self-assembling peptide nanofibres.
Figure 2: Fluorescent βTail fusion proteins stably integrate into Q11 nanofibres and microgels in smoothly gradated amounts, alone or in combination, without loss of activity.
Figure 3: TEM, circular dichroism and fluorescence measurements confirmed the rapid co-assembly of βTail polypeptides into Q11 nanofibres.
Figure 4: The βTail platform is capable of integrating an enzyme into nanofibres in precise amounts, without loss of enzyme activity.
Figure 5: Nanofibres exhibiting βTail fusion proteins raised antibody responses in mice that could be adjusted by changing the protein content.


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This research was supported by the National Institutes of Health (NIBIB, 1R01EB009701; NCI, U54 CA151880; NIAID, 1F32AI096769 and 5R21AI09444), the Chicago Biomedical Consortium with support from the Searle Funds at the Chicago Community Trust, and the National Science Foundation (CHE-0802286). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Biomedical Imaging and BioEngineering, the National Institute of Allergy and Infectious Disease, the National Cancer Institute, or the National Institutes of Health.

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Authors and Affiliations



J.H.C. directed the work, designed experiments, analysed the data and wrote the paper. G.A.H. designed experiments, conducted experiments, analysed the data and wrote the paper. T.S., J.Z.G., Y.F.T. and H.H. performed experiments and analysed the results. A.S.C. designed experiments and analysed data.

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Correspondence to Joel H. Collier.

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Competing interests

J.H.C. and G.A.H. are named as inventors on a patent application filed by the University of Chicago (US Patent Application No. 14/209,757, ‘Methods and compositions involving fibrillizing polypeptides for nanofibers’) that covers the technology described in this paper.

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Hudalla, G., Sun, T., Gasiorowski, J. et al. Gradated assembly of multiple proteins into supramolecular nanomaterials. Nature Mater 13, 829–836 (2014).

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