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We present a new method for the surface capture of proteins in cell-free protein synthesis (CFPS). We demonstrate the spontaneous self-assembly of the protein BslA into functionalizable surfaces on the surface of a CFPS reaction chamber. We show that proteins can be covalently captured by such surfaces, using "Catcher/Tag" technology. Importantly, proteins of interest can be captured either when synthesised by CFPS above the BslA surfaces, or when added as pure protein. The simplicity and cost efficiency of this method suggest that it will find many applications in cell-free-based methods.
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http://dx.doi.org/10.3389/fbioe.2022.915035 | DOI Listing |
Biotechnol Biotechnol Equip
February 2025
Department of Bioengineering, University of Washington, Seattle, WA, USA.
Phage-like particles (PLPs) are fabricated self-assembling nanoparticles derived from the structural elements of bacteriophages. These particles have biotechnological utility because of the ability to easily modify surface chemistry and compartmentalize nucleic acids or other materials. A consequential implementation of PLPs in diagnostics is as process controls in nucleic acid amplification tests, where control RNAs are packaged within the protein capsid and protected from degradation by RNases in the sample matrix.
View Article and Find Full Text PDFNanoscale Adv
August 2025
Department of Chemistry, Birla Institute of Technology and Science Pilani Hyderabad Campus, Jawahar Nagar Hyderabad 500 078 Telangana India
Amyloid nanostructures are gaining attention as bio-compatible materials with diverse potential applications. The formation/self-assembly of amyloid fibrils, though implicated in the prognosis of several neurodegenerative diseases, contrastingly can also be explored for their functional properties owing to their unique stability and strength. The physicochemical environment and amino-acid composition are characteristic and specifically crucial for a protein/peptide to form amyloid fibrils.
View Article and Find Full Text PDFbioRxiv
August 2025
Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
Self-assembling protein nanoparticles are being increasingly utilized in the design of next-generation vaccines due to their ability to induce antibody responses of superior magnitude, breadth, and durability. Computational protein design offers a route to novel nanoparticle scaffolds with structural and biochemical features tailored to specific vaccine applications. Although strategies for designing new self-assembling proteins have been established, the recent development of powerful machine learning-based tools for protein structure prediction and design provides an opportunity to overcome several of their limitations.
View Article and Find Full Text PDFBiomacromolecules
August 2025
Department of Chemical Engineering, University of Massachusetts Amherst, Amherst, Massachusetts 01003, United States.
Complex coacervation has emerged as a powerful model for studying the self-assembly of intrinsically disordered proteins (IDPs) in biological condensates in cells. We characterized the phase behavior and rheology of coacervates formed from peptides with regular repeating sequences to examine the effects of charge patterning and hydrophobicity on coacervate stability and material properties. Our results show that increasing the size of charged blocks enhances salt resistance via electrostatic cooperativity, while incorporating small hydrophobic segments further stabilizes coacervates and increases viscosity through hydrophobic clustering.
View Article and Find Full Text PDFAppl Biochem Biotechnol
August 2025
Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, 04510, Mexico.
Protein nanocages are a group of compartments naturally enclosing nucleic acids or proteins for biological purposes. Such materials have also inspired the design of novel proteins displaying self-assembling properties. The most studied protein nanocages are viral capsids and their derivative virus-like particles (VLPs), which consist of any or all of the structural proteins of the virion but lack nucleic acids and are therefore non-infectious.
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