Development of Methods for the Analysis of Feline-Interferon Omega Activity, Quantity and Quality
Wen-Hua Lin
Abstract
Type I Interferons (IFNs) are well known cytokines that function as key components of the host immune response with broad and strong antiviral, antitumor, and immunomodulatory effects. The first animal interferon product was developed by Toray Industries Inc. of Japan in 1994 using nucleopolyhedrovirus (NPV) as a carrier to treat feline calicivirus. Since market demand for interferon products for the treatment of animals will increase in the future, we used Feline-Interferon Omega (FeIFN-ω) as a proof-of-concept example for the establishment of interferon analysis techniques and protocols. To test for interferon activity, vesicular stomatitis virus was added to Felis catus 9 (Fc9) cell lines induced with FeIFN-ω, . The interferon suppressed viral growth in the cell lines and interferon activity was calculated based on half of the maximal effective dose (EC50). For the calculated EC50, the relative standard deviation of repeatability was 6.39% and the relative standard deviation of accuracy was 3.18%, which meet the standards for biological validation. In addition, we developed a high-performance liquid chromatography analytical method for the determination of was established for the quantification of inteferons as well as for the determination of sample contents (closely related protein variants, degradation products, and other high-molecular weight substances). In this method, samples and standards are diluted and filtered by deionized water, loaded onto a Vydac® C4 (5μm, inner diameter 4.6×250mm) liquid chromatography column, with a 0.1% trifluoroacetic acid/90% acetonitrile solution used as a mobile phase gradient, and a photodiode array detector was used for detection at a wavelength of 214 nm and a flow rate of 0.5mL/min. The linear range for the analysis is 14 - 72μg/mL, (R2 = 0.9952), while the relative standard deviation of repeatability and the relative standard deviation of precision were 1.91% and2.92%, respectively, which meet the standards for analytical validation.
Establishment of ornamental KHV qPCR and monitor result
Yu-Ting Liu
Abstract
Animal health research institute assists stationed aquarium companies in Pingtung Agricultural Biotechnology Park and their satellite fish farm to monitor diseases, and establish model of bio-safety. Furthermore, leading in management of health and the conception of animal health and medicine usage. Our lab set up methods of diseases examination vigorously, one of them is Koi herpes virus (KHV). The main host of KHV is cyprinidae, one of the most important pathogen. Therefore, we consulted the paper that OIE refers, establish and optimize the method of KHV qPCR by adjusting reagent formula and temperature of qPCR program. Drawing on KHV detection in fish farms, and assist owners of fish farms implement the management of aquarium quarantine. Connecting with traceability and providing with international competitiveness.