Veterinary Research Forum
Vet Res Forum

A methodological approach for production and purification of polyclonal ‎antibody against dog IgG ‎

Document Type : Original Article

Authors

Somayeh Sadeghi 1, 2
Leili Aghebati-Maleki 1, 2
Samira Nozari 3
Jafar Majidi 1, 2

1 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

2 Department of Immunology, Faculty of Medicine, Tabriz ‎University of Medical Sciences, Tabriz, Iran

3 Department of Immunology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

Abstract
Antibodies are a class of biomolecules that has an important role in the immune system and lots of applications in biotechnological methods and in pharmaceutics. Production and purification of antibodies in laboratory animals is one of the first ways to manufacture of these prominent tools. The obtained antibodies from these process could be used in various types of bioassay techniques such as enzyme linked immunosorbent assay (ELISA), radioimmunoassay, etc. Also, antibodies employed in diagnostics applications in humans and other animals in order to detect specific antigens.In this study, we aimed to produce and purify anti-dog IgG via immunizing rabbits with dog IgG in combination with Freund's adjuvant. Polyclonal IgG were purified by ion exchange chromatography and then the purified antibody was labeled with horse radish peroxidase (HPR). Direct ELISA was used to determine the optimum titer and cross-reactivity of HRP conjugated IgG. The purity of various IgG preparations and the optimum dilution of prepared HRP conjugated IgG, respectively, was about 95.00% and 1:8000. This study showed that efficiency ion-exchange chromatography could be an appropriate method for purification of IgG antibodies. This antibody could be a useful tool for future dog immune diagnosis tests. This product characterization shown here sets the foundations for future work on dog IgGs.

Keywords

Anti-dog IgG
‎ HRP-conjugation
‎ Ion-exchange chromatography
‎ Polyclonal antibody

Subjects

  1.  

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Veterinary Research Forum
Volume 9, Issue 1
Winter 2018
Pages 13-18

  • Receive Date 19 February 2017
  • Revise Date 10 July 2017
  • Accept Date 11 July 2017

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