Volume 36, Issue 4 (2-2013)                   Research in Medicine 2013, 36(4): 167-172 | Back to browse issues page

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Molecular and Cellular Biology Department, Royan Institute, Isfahan, Iran , agholami@pasteur.ac.ir|
Abstract:   (8455 Views)
Abstract Background: Rabies is the most important zoonosis in country. Anti-viral antibody conjugated with Fluorescein Isothiocyanate (FITC), is used for diagnosis of Rabies. We import this reagent from abroad. In this study we aimed to prepare this conjugated fluorescein antibody and assess its utility. Materials and methods: Rabbits were immunized with rabies vaccine, then serum samples were taken and concentrated by ammonium sulfate and treated by gel-filtration. To purify IgG concentrated serum protein ion exchange chromatography was done. Validation method such as Dot-blot technique was applied for confirmation of the extracted IgG and for its specificity. Anti rabies antibody was conjugated with FITC and evaluated against commercial conjugate. Potency of locally made antibody was considered acceptable if this antibody had enough specificity and brightness in comparison to commercial antibody. Results: Maximum quantity of IgG was extracted at 100 mM NaCl in ion exchange chromatography. Comparison of two fractions showed better efficacy of conjugation with 100 mM NaCl comparing with 200mM NaCl fraction. Conclusion: The study showed the feasibility of preparation of the fluorescent antibody with comparable efficacy to the commercial antibody. Keywords: Fluorescein Isothiocyanate, Immunofluorescent test, Gel filtration, Dot blotting, Ion Exchange Chromatography.
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