Ff-nano, short functionalized nanorods derived from Ff (f1, fd, or M13) filamentous bacteriophage

Abstract

F-specific filamentous phage of Escherichia coli (Ff: f1, M13, or fd) are long thin filaments (860 nm × 6 nm). They have been a major workhorse in display technologies and bionanotechnology; however, some applications are limited by the high length-to-diameter ratio of Ff. Furthermore, use of functionalized Ff outside of laboratory containment is in part hampered by the fact that they are genetically modified viruses. We have now developed a system for production and purification of very short functionalized Ff-phage-derived nanorods, named Ff-nano, that are only 50 nm in length. In contrast to standard Ff-derived vectors that replicate in E. coli and contain antibiotic-resistance genes, Ff-nano are protein-DNA complexes that cannot replicate on their own and do not contain any coding sequences. These nanorods show an increased resistance to heating at 70∘C in 1% SDS in comparison to the full-length Ff phage of the same coat composition. We demonstrate that functionalized Ff-nano particles are suitable for application as detection particles in sensitive and quantitative “dipstick” lateral flow diagnostic assay for human plasma fibronectin.

Arming Filamentous Bacteriophage, a Nature-Made Nanoparticle, for New Vaccine and Immunotherapeutic Strategies

“…Bacteriophages can be inactivated without changing their immunogenicity; phage-based vaccines formulated to include heat-inactivated or UV-inactivated virions might represent safer alternatives to live phage vaccines [118,119]. Shorter (50 nm × 6 nm) pIII-recombinant Ff-derived particles (fd-nano) that do not carry any viral gene or antibiotic resistance and cannot replicate within bacterial cells or integrate into the bacterial genome have also been proposed [120].…”

Evaluation of Three Morphologically Distinct Virus-Like Particles as Nanocarriers for Convection-Enhanced Drug Delivery to Glioblastoma

“…MS2 and TMV proteins were expressed and purified using previously published methods [15,28,30]. Nanophage was expressed and purified using an adapted method from a previously published report [35]. All proteins were purified using anion exchange chromatography with a diethylaminoethanol (DEAE) Sepharose column followed by size exclusion chromatography.…”

“…The last member of the VLP panel was a nanoscale variation of the filamentous fd phage. The nanophage (NP) VLP was recently reported as a new potential nanocarrier [35] but has not previously been functionalized and evaluated for its drug delivery potential. This panel therefore encompasses disparate VLP morphologies, while maintaining relatively consistent dimensions of 15–50 nm.…”

Phage‐Enabled Nanomedicine: From Probes to Therapeutics in Precision Medicine

“…Phages are a powerful building material and can be assembled into highly organized nanomaterials with various desirable characteristics . In addition to using the natural structure of phage itself as a nanomaterial, phages can easily be genetically engineered to display peptides that provide affinity characteristics and serve as a biointeractive peptide motif, or even to form self‐assembled nanomaterials .…”