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Rapid fabrication of precise high-throughput filters from membrane protein nanosheets

Nature Materials volume 19, pages 347–354 (2020)Cite this article

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Abstract

Biological membranes are ideal for separations as they provide high permeability while maintaining high solute selectivity due to the presence of specialized membrane protein (MP) channels. However, successful integration of MPs into manufactured membranes has remained a significant challenge. Here, we demonstrate a two-hour organic solvent method to develop 2D crystals and nanosheets of highly packed pore-forming MPs in block copolymers (BCPs). We then integrate these hybrid materials into scalable MP-BCP biomimetic membranes. These MP-BCP nanosheet membranes maintain the molecular selectivity of the three types of β-barrel MP channels used, with pore sizes of 0.8 nm, 1.3 nm, and 1.5 nm. These biomimetic membranes demonstrate water permeability that is 20–1,000 times greater than that of commercial membranes and 1.5–45 times greater than that of the latest research membranes with comparable molecular exclusion ratings. This approach could provide high performance alternatives in the challenging sub-nanometre to few-nanometre size range.

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Fig. 1: Stable β-barrel MP-BCP-based scalable membranes.
Fig. 2: 2D crystals or nanosheets of three β-barrel channel proteins reconstituted in BCP membrane matrices.
Fig. 3: 2D OmpF nanosheets can be assembled on a PC (50 nm), PES (MP005) or aluminium oxide (0.02 μm Anodisc) substrate.
Fig. 4: Three scalable biomimetic membranes based on β-barrel channel MPs and BCPs demonstrate distinct molecular separations and enhanced pure-water permeability compared with current commercial membranes.
Fig. 5: Comparison of water permeability (LMH bar–1) and MWCO (Da) of MP-based membranes with commercial NF or ultrafiltration membranes tested in the same experimental setup.

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The data supporting the findings of this study are available within the article and its supplementary information files and available from the authors upon reasonable request.

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Acknowledgements

The authors acknowledge financial support from the National Science Foundation (NSF) CAREER grant (CBET-1552571), NSF grant CBET-1709522 and NSF grant CBET-1804836 to M.K. for this work. T.C. and E.D.G. acknowledge financial support from NSF DMR-1609417. The authors also thank M. Hazen and J. Cantolina for their help with cross-sectional sample preparation. We thank L. Movileanu for the kind gift of the plasmid for expressing the FhuA ΔC/Δ4L protein.

Author information

Author notes

  1. Yu-Ming Tu, Woochul Song & Laxmicharan Samineni

    Present address: Department of Chemical Engineering, The University of Texas at Austin, Austin, TX, USA

  2. Manish Kumar

    Present address: Department of Civil, Architectural and Environmental Engineering, The University of Texas at Austin, Austin, TX, USA

  3. These authors contributed equally: Yu-Ming Tu, Woochul Song, Tingwei Ren.

Authors and Affiliations

  1. Department of Chemical Engineering, Pennsylvania State University, University Park, PA, USA

    Yu-Ming Tu, Woochul Song, Tingwei Ren, Ratul Chowdhury, Tyler E. Culp, Laxmicharan Samineni, Chao Lang, Drew Carson, Yuxuan Dai, Miaoci Zhang, Enrique D. Gomez & Manish Kumar

  2. Department of Civil, Environmental, & Construction Engineering, Texas Tech University, Lubbock, TX, USA

    Yue-xiao Shen

  3. Department of Chemistry, University of Kentucky, Lexington, KY, USA

    Prasangi Rajapaksha & Yinai Wei

  4. Department of Materials Science and Engineering, The Pennsylvania State University, University Park, PA, USA

    Chao Lang & Robert J. Hickey

  5. Department of Chemical and Biochemical Engineering, Rutgers University, Piscataway, NJ, USA

    Alina Thokkadam

  6. Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA, USA

    Arwa Mukthar

  7. Applied Biomimetic, Gaithersburg, MD, USA

    Andrey Parshin & Mariusz Grzelakowski

  8. Department of Biomedical Engineering, Pennsylvania State University, University Park, PA, USA

    Janna N. Sloand & Scott H. Medina

  9. Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY, USA

    Dibakar Bhattacharya

  10. Department of Chemical and Biomolecular Engineering, University of Notre Dame, Notre Dame, IN, USA

    William A. Phillip

  11. Materials Research Institute, Pennsylvania State University, University Park, PA, USA

    Robert J. Hickey & Manish Kumar

  12. Department of Civil and Environmental Engineering, Pennsylvania State University, University Park, PA, USA

    Manish Kumar

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Contributions

Y.-M.T., W.S., T.R. and M.K. conceived and designed the research. T.R., W.S., Y.-M.T., Y.-x.S. P.R., T.E.C. and L.S. performed the experiments, with the assistance of R.C., C.L., A.T., D.C., Y.D., A.M. and M.Z. in specialized analytic tools. T.R., Y.-M.T., P.R., A.P., J.N.S., S.H.M. and M.G. contributed to the protein production. Y.-M.T., W.S., T.R., R.C., T.E.C., L.S., D.B., W.A.P., E.D.G., R.J.H., Y.W. and M.K. analysed and interpreted the data and results. T.R., W.S., Y.-M.T. and M.K. co-wrote the paper.

Corresponding author

Correspondence to Manish Kumar.

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Competing interests

A.P. and M.G. are employed by Applied Biomimetic, which aims to commercialize biomimetic membranes similar to those presented in this Article.

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Supplementary discussion, Tables 1–5, Figures 1–19, references 1–31.

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Tu, YM., Song, W., Ren, T. et al. Rapid fabrication of precise high-throughput filters from membrane protein nanosheets. Nat. Mater. 19, 347–354 (2020). https://doi.org/10.1038/s41563-019-0577-z

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