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  • Construction and physiochemical characterisation of a multi-composite, potential oral vaccine delivery system (VDS).

Construction and physiochemical characterisation of a multi-composite, potential oral vaccine delivery system (VDS).

International journal of pharmaceutics (2014-04-01)
Marie W Pettit, Paul D R Dyer, John C Mitchell, Peter C Griffiths, Bruce Alexander, Beatrice Cattoz, Richard K Heenan, Stephen M King, Ralf Schweins, Frank Pullen, Stephen R Wicks, Simon C W Richardson
ABSTRACT

An increasing human population requires a secure food supply and a cost effective, oral vaccine delivery system for livestock would help facilitate this end. Recombinant antigen adsorbed onto silica beads and coated with myristic acid, was released (∼15% (w/v)) over 24 h at pH 8.8. At pH 2, the myristic acid acted as an enteric coating, protecting the antigen from a variety of proteases. The antigen adsorbed onto silica particles, coated in myristic acid had a conserved secondary structure (measured by circular dichroism (CD) spectroscopy) following its pH-triggered release. Small angle neutron scattering (SANS) was used to measure the thickness of the adsorbed antigen, finding that its adsorbed conformation was slightly greater than its solution radius of gyration, i.e. 120-160 Å. The addition of myristic acid led to a further increase in particle size, with scattering data consistent with an acid thickness slightly greater than a monolayer of fully extended alkyl chains and a degree of hydration of around 50%. Whilst adsorbed onto the silica and coated in myristic acid, the protein was stable over 14 days at 42 °C, indicating a reduced need for cold chain storage. These data indicate that further investigation is warranted into the development of this technology.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ammonium acetate, BioUltra, ≥99.0%
Supelco
Myristic acid, analytical standard
Sigma-Aldrich
Acrylamide, for Northern and Southern blotting, powder blend
Sigma-Aldrich
Glycerol solution, 83.5-89.5% (T)
Supelco
Sodium hydroxide solution, 49-51% in water, eluent for IC
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Sodium hydroxide, BioUltra, suitable for luminescence, ≥98.0% (T), pellets
Sigma-Aldrich
2-Mercaptoethanol, BioUltra, Molecular Biology, ≥99.0% (GC)
Supelco
Lithium chloride solution, 1 M in ethanol
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Acetone, analytical standard
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Sodium bicarbonate, tested according to Ph. Eur.
Sigma-Aldrich
Sodium azide, purum p.a., ≥99.0% (T)
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Potassium acetate solution, BioUltra, Molecular Biology, 5 M in H2O
Sigma-Aldrich
Sodium azide, BioUltra, ≥99.5% (T)
Sigma-Aldrich
Sodium hydroxide solution, BioUltra, Molecular Biology, 10 M in H2O
Sigma-Aldrich
Lithium chloride, BioUltra, Molecular Biology, anhydrous, ≥99.0% (AT)
Supelco
Ammonium acetate, LiChropur, eluent additive for LC-MS
Sigma-Aldrich
Acrylamide, purum, ≥98.0% (GC)
Sigma-Aldrich
Myristic acid, ≥95%, FCC, FG
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Myristic acid, natural, ≥98.5%, FG
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Ammonium acetate solution, BioUltra, ~5 M in H2O
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Myristic acid, ≥98.0% (GC)
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3-Ethyl-2,4-pentanedione, mixture of tautomers, 98%
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Acrylamide, Molecular Biology, ≥99% (HPLC)
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2-Mercaptoethanol, ≥99.0%
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99% (HPLC), powder
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Sodium bicarbonate, BioXtra, 99.5-100.5%
Sigma-Aldrich
Sodium hydroxide solution, 1.0 N, BioReagent, suitable for cell culture
Sigma-Aldrich
Ammonium acetate, Molecular Biology, ≥98%
Sigma-Aldrich
Sodium azide, ReagentPlus®, ≥99.5%