Application of filamentous phages

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Application of filamentous phages In
nanobiotechnology
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1) Peptide phage display isolate binders of
semiconductors
Phage structure and phage display selection
process. (a) Schematic diagram of phage and its
genome and (b) phage-display process to identify
specific binding peptide motifs against desired
targets.
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2) Use the isolated phages as nucleation centers
for the fabrication of nanoparticles and
nanowires
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Application of filamentous phages As nanomedicines
Phages as vehicles for gene and vaccine delivery
Phages as tool for imaging
Phages as vehicles for gene and drug delivery
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Combine 1) Gene delivery 2) Specific
targeting 3) Imaging
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Components of a targeted drug carrying platform
Potency
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The filamentous bacteriophage as a targeted
drug-carrying nanomedicine
6nm
800-2000 nm (dependent on the size of the
packaged genome)
Scheme of the filamentous Fd phage. In out
system, the minor coat protein, p3 (g3p) carries
the targeting moiety which the drug, and the
engineered release mechanism are on p8 (g8p).
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IgG complexed to fUSE5-ZZ phage through a
p3-displayed ZZ domain
A
B C
fUSE5-ZZ phage used for targeting. A Scheme of
the filamentous fUSE5-ZZ phage. B. Evaluationof
ZZ domain display by an immunoblot. Phage
particles (each lane is identified with the
corresponding phage name below it)were separated
by SDS/PAGE and electro-botted onto
nitrocellulose, and the p3 minor coat protein or
the derived ZZ domain-p3 fused derivative was
detected with an anti-p3 antibody. The upper
arrow marks the position of the ZZ-p3 fusion,
while the lower arrow marks the position of the
wild-type p3 coat protein. C. Evaluation of
antibody binding capacity by competitive ELISA.
1012 fUSE5-ZZ phages were complexed with 0.6 mg
(3 dil) or 0.2 mg (9 dil) of HRP-conjugated
rabbit anti mouse IgG as tracer, in the presence
of varying concentrations of protein-A purified
human IgG. The residual HRP on the phages was
detected using the substrate TMB.
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Preparation of drug-linker adduct Chloramphenicol
was modified in two steps to create an ester bond
between CAM and a linker (originated in glutatic
anhydride) The linker CAM complex is activated
for lysine conjugation by the NHS procedure.
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Kinetics of drug release by serum
esterases (here 10 horse serum analysis by HPLC)
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Partial growth inhibition of staphylococcus
aureus by antibody-targeted drug-carrying phages
(3000 Cam/phage)
Similar Partial growth inhibition was obtained by
peptide-targeted phages
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• The limitations
• Limited arming efficiency due to drug
  hydrophobicity
• Solubility of the platform also affected
• Vulnerability of the targeting moiety (ZZ domain)
  to amine chemistry