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Biochemical and aerosol characterization of ricin for use in non‐clinical efficacy studies
Author(s) -
Barnewall Roy E.,
Riffle Carol G.,
Jones Randy L.,
Guistino David J.,
Chou Richard M.,
Anderson Mike S.,
Vassar Michelle L.,
Howland Carrie A.
Publication year - 2017
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.21980
Subject(s) - ricin , chemistry , aerosolization , bioassay , potency , chromatography , toxicity , pharmacology , toxin , reticulocyte , lysis , lethal dose , sodium dodecyl sulfate , median lethal dose , biochemistry , in vitro , biology , medicine , inhalation , anesthesia , organic chemistry , messenger rna , gene , genetics
Ricin toxin may be used as a biological warfare agent and no medical countermeasures are currently available. Here, a well‐characterized lot of ricin was aerosolized to determine the delivered dose for future pre‐clinical efficacy studies. Mouse intraperitoneal (IP) median lethal dose (LD 50 ) bioassay measured potency at 5.62 and 7.35 μg/kg on Days 0 and 365, respectively. Additional analyses included total protein, sodium dodecyl sulfate polyacrylamide gel electrophoresis, Western blotting, and rabbit reticulocyte lysate activity assay. The nebulizer aerosol produced consistent concentrations (2.5 × 10 3 , 5.0 × 10 3 , 1.0 × 10 4 , and 1.5 × 10 4 μg/mL) and spray factor values. The aerosol particle size distribution was of sufficient size to deposit in lung alveoli (1.12–1.43 μm). Ricinus communi s Agglutinin II (RCA 60), prepared at 19 mg/mL in phosphate‐buffered saline, pH 7.8, and stored at −70°C, maintained attributes for toxicity following 1‐year storage and aerosolized consistently.