Abstract: Various nanomaterials can be used as a drug delivery
vehicles in nanomedicine, called nanocarriers. They can either be
organic or inorganic, synthetic or natural-based. Although synthetic
nanocarriers are easier to produce, they can often be toxic for the
organism and thus not suitable for use in treatment. From naturalbased
nanocarriers, the most commonly used are protein cages or
viral capsids. In this work, virus bacteriophage λ was used for
delivery of different cytotoxic drugs (cisplatin, carboplatin,
oxaliplatin and doxorubicin). Large quantities of phage λ were
obtained from phage λ-producing strain of E. coli cultivated in
medium with 0.2% maltose. After killing of E. coli with chloroform
and its removal by centrifugation, the phage was concentrated by
ultracentrifugation at 130 000×g and 4°C for 3 h. The encapsulation
of the drugs was performed by infusion method and four different
concentrations of the drugs were encapsulated (200; 100; 50; 25
μg·mL-1). Free drug molecules were removed by filtration. The
encapsulation was verified using the absorbance for doxorubicin and
atomic absorption spectrometry for platinum cytostatics. The amount
of encapsulated drug linearly increased with the increasing
concentration of applied drug with the determination coefficient
R2=0.989 for doxorubicin; R2=0.967 for cisplatin; R2=0.989 for
carboplatin and R2=0.996 for oxaliplatin. The overall encapsulation
efficiency was calculated as 50% for doxorubicin; 8% for cisplatin;
6% for carboplatin and 10% for oxaliplatin.
Abstract: To ensure targeting of apoferritin nanocarrier with
encapsulated doxorubicin drug, we used a peptide linker based on a
protein G with N-terminus affinity towards Fc region of antibodies.
To connect the peptide to the surface of apoferritin, the C-terminus of
peptide was made of cysteine with affinity to gold. The surface of
apoferritin with encapsulated doxorubicin (APODOX) was coated
either with gold nanoparticles (APODOX-Nano) or gold(III) chloride
hydrate reduced with sodium borohydride (APODOX-HAu). The
reduction with sodium borohydride caused a loss of doxorubicin
fluorescent properties and probably accompanied with the loss of its
biological activity. Fluorescent properties of APODOX-Nano were
similar to the unmodified APODOX; therefore it was more suited for
the intended use. To evaluate the specificity of apoferritin modified
with antibodies, ELISA-like method was used with the surface of
microtitration plate wells coated by the antigen (goat anti-human IgG
antibodies). To these wells, the nanocarrier was applied. APODOX
without the modification showed 5× lower affinity to the antigen than
APODOX-Nano modified gold and targeting antibodies (human IgG
antibodies).