Merck
  • Home
  • Search Results
  • Effective multiple oral administration of reverse genetics engineered infectious bursal disease virus in mice in the presence of neutralizing antibodies.

Effective multiple oral administration of reverse genetics engineered infectious bursal disease virus in mice in the presence of neutralizing antibodies.

The journal of gene medicine (2015-05-02)
Ákos Hornyák, Kai S Lipinski, Tamás Bakonyi, Petra Forgách, Ernő Horváth, Attila Farsang, Susan J Hedley, Vilmos Palya, Tibor Bakács, Imre Kovesdi
ABSTRACT

Despite spectacular successes in hepatitis B and C therapies, severe hepatic impairment is still a major treatment problem. The clinically tested infectious bursal disease virus (IBDV) superinfection therapy promises an innovative, interferon-free solution to this great unmet need, provided that a consistent manufacturing process preventing mutations or reversions to virulent strains is obtained. To address safety concerns, a tissue culture adapted IBDV vaccine strain V903/78 was cloned into cDNA plasmids ensuring reproducible production of a reverse engineered virus R903/78. The therapeutic drug candidate was characterized by immunocytochemistry assay, virus particle determination and immunoblot analysis. The biodistribution and potential immunogenicity of the IBDV agent was determined in mice, which is not a natural host of this virus, by quantitative detection of IBDV RNA by a quantitative reverse transcriptase-polymerase chain reaction and virus neutralization test, respectively. Several human cell lines supported IBDV propagation in the absence of visible cytopathic effect. The virus was stable from pH 8 to pH 6 and demonstrated significant resistance to low pH and also proved to be highly resistant to high temperatures. No pathological effects were observed in mice. Single and multiple oral administration of IBDV elicited antibodies with neutralizing activities in vitro. Repeat oral administration of R903/78 was successful despite the presence of neutralizing antibodies. Single oral and intravenous administration indicated that IBDV does not replicate in mammalian liver alleviating some safety related concerns. These data supports the development of an orally delivered anti-hepatitis B virus/ anti-hepatitis C virus viral agent for human use.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Phenol, BioXtra, ≥99.5% (GC)
Sigma-Aldrich
Phenol solution, Saturated with 0.1 M citrate buffer, pH 4.3 ± 0.2, BioReagent, for molecular biology
Sigma-Aldrich
Phenol solution, Equilibrated with 10 mM Tris HCl, pH 8.0, 1 mM EDTA, BioReagent, for molecular biology
Sigma-Aldrich
Phenol, puriss., meets analytical specification of Ph. Eur., BP, USP, ≥99.5% (GC), crystalline (detached)
Sigma-Aldrich
Phenol, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.5-100.5% (GC)
Sigma-Aldrich
Phenol, puriss. p.a., ACS reagent, reag. Ph. Eur., 99.0-100.5%
Sigma-Aldrich
Phenol, BioUltra, for molecular biology, TE-saturated, ~73% (T)
Sigma-Aldrich
Phenol, ≥96.0% (calc. on dry substance, T)
Sigma-Aldrich
Phenol, natural, 97%, FG
Sigma-Aldrich
Phenol, contains hypophosphorous as stabilizer, loose crystals, ACS reagent, ≥99.0%
Sigma-Aldrich
Phenol, ≥99%
Sigma-Aldrich
Phenol, unstabilized, ReagentPlus®, ≥99%
Sigma-Aldrich
Phenol, for molecular biology
Sigma-Aldrich
Ethidium bromide, BioReagent, for molecular biology, powder
Sigma-Aldrich
Ethidium bromide, ~95% (HPLC)
Sigma-Aldrich
Phenol, unstabilized, purified by redistillation, ≥99%
Sigma-Aldrich
Liquified Phenol, ≥89.0%
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 10 mg/mL in H2O
Sigma-Aldrich
Ethidium bromide solution, BioReagent, for molecular biology, 500 μg/mL in H2O
Supelco
Pyridaben, PESTANAL®, analytical standard
Sigma-Aldrich
Ethidium bromide solution, for fluorescence, ~1% in H2O
Sigma-Aldrich
Chloroform, anhydrous, contains amylenes as stabilizer, ≥99%
Sigma-Aldrich
Chloroform, anhydrous, ≥99%, contains 0.5-1.0% ethanol as stabilizer
Sigma-Aldrich
Chloroform, ≥99%, PCR Reagent, contains amylenes as stabilizer