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  • Characterization of biomolecular nanoconjugates by high-throughput delivery and spectroscopic difference.

Characterization of biomolecular nanoconjugates by high-throughput delivery and spectroscopic difference.

Nanomedicine (London, England) (2012-09-05)
Robert K Delong, Azure Risor, Masaaki Kanomata, Amanda Laymon, Brooke Jones, Scott D Zimmerman, Joseph Williams, Colette Witkowski, Mathew Warner, Michael Ruff, Richard Garrad, John K Fallon, Anthony J Hickey, Reza Sedaghat-Herati
ABSTRACT

Nanoparticle conjugates have the potential for delivering siRNA, splice-shifting oligomers or nucleic acid vaccines, and can be applicable to anticancer therapeutics. This article compares tripartite conjugates with gold nanoparticles or synthetic methoxypoly(ethylene glycol)-block-polyamidoamine dendrimers. Interactions with model liposomes of a 1:1 molar ratio of tripalmitin:cholesterol or phospholipid:cholesterol were investigated by high-throughput absorbance, as well as fluorescence difference and cellular luminescence assays. Spectral differences and dynamic light-scattering spectroscopy shifts demonstrated the interaction of conjugates with liposomes. Biological activity was demonstrated by upregulation of gene expression via splice-shifting oligomers, delivery of anti-B-Raf siRNA in cultured human cancer cells or tuberculosis antigen 85B plasmid expression vector in a coculture model of antigen presentation. The data suggests that gold nanoparticles and methoxypoly(ethylene glycol)-block-polyamidoamine dendrimer nanoconjugates may have potential for binding, stabilization and delivery of splice-shifting oligomers, siRNA and nucleic acid vaccines for preclinical trials.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Poly(ethylene glycol) methyl ether, average Mn 550
Sigma-Aldrich
Poly(ethylene glycol) methyl ether, average Mn ~2,000
Sigma-Aldrich
Poly(ethylene glycol) methyl ether, average Mn 750
Sigma-Aldrich
Poly(ethylene glycol) methyl ether, BioUltra, 500
Sigma-Aldrich
Poly(ethylene glycol) methyl ether, average Mn 5,000
Sigma-Aldrich
Poly(ethylene glycol) methyl ether, average Mn 10,000
Sigma-Aldrich
Poly(ethylene glycol) methyl ether, average Mn 20,000
Sigma-Aldrich
Methoxypolyethylene glycol 350, average mol wt 350