Establishment of an Antiviral Activity Assay and the Identification and Partial Purification of Interferon-like Protein from Rainbow Trout Gonadal Cells (RTG-2)
Shih-Bin Lin, Chih-Hsin Lin and Ya-Li Hsu (2001) In this study, a rapid (performed within 24 h) and effective bioassay method free from overestimation or false positives for antiviral activity due to virus residue (within an MOI range of 6.4 × 10-4 to 0.4) was established. Antiviral proteins, generated by infectious pancreatic necrosis virus (IPNV) induction of rainbow trout gonadal cells (RTG-2), were partially purified by HPLC size exclusion chromatography and native polyacrylamide gel electrophoresis (PAGE); the chromatography demonstrated 2 possible antiviral proteins with molecular weights of 100 and 18 kDa, respectively. Electrophoresis produced 1 antiviral protein with a molecular weight of about 100 kDa identified through Western blotting with polyclonal anti-human IFN-γ. The yield of antiviral protein obtained from 10-dold cells was 1.6-3.2 times higher than that from 6-d-old cells.
Keywords
Interferon-like protein, Antiviral protein, RTG-2, IPNV.
