Volume 36, Issue 4 (2-2013)
Research in Medicine 2013, 36(4): 167-172 |
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Gholami Arjenak V, Mozhgani S H R M, Miandeh, N, Janani A R, Ajorloo M, Zavareh A R, et al . Production of rabies fluorescent antibody conjugated and determination of its function. Research in Medicine 2013; 36 (4) :167-172
URL: http://pejouhesh.sbmu.ac.ir/article-1-1088-en.html
URL: http://pejouhesh.sbmu.ac.ir/article-1-1088-en.html
Vahidi Gholami Arjenak 
, Seyed Hamid Reza Miandeh Mozhgani , Narges Miandeh, , Ali Reza Janani , Mehdi Ajorloo , Ali reza Zavareh , Seyed Mohamad Atyabi , Nabiallah Namvar Asl , Saeed Jodairi Eslami , Alireza Gholami
, Seyed Hamid Reza Miandeh Mozhgani , Narges Miandeh, , Ali Reza Janani , Mehdi Ajorloo , Ali reza Zavareh , Seyed Mohamad Atyabi , Nabiallah Namvar Asl , Saeed Jodairi Eslami , Alireza Gholami
Molecular and Cellular Biology Department, Royan Institute, Isfahan, Iran , agholami@pasteur.ac.ir|
Abstract: (11276 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.
Keywords: Fluorescein Isothiocyanate, Immunofluorescent test, Gel filtration, Dot blotting, Ion Exchange Chromatography.
Type of Study: Original |
Subject:
Interdisciplinary (Educational Management, Educational research, Statistics, Medical education
Received: 2013/03/16 | Published: 2013/02/15
Received: 2013/03/16 | Published: 2013/02/15
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This Journal is licensed under a Creative Commons Attribution 4.0 International License | Research in Medicine
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