Development of DNA-Designed Avian IgY Antibodies for Quantitative Determination of Bovine Interferon-Gamma |
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Authors: | Gholamreza Nikbakht Brujeni Sayed Amir Hossein Jalali Mohammad Kazem Koohi |
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Institution: | (1) Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran;(2) Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran;(3) Department of Toxicology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran |
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Abstract: | Interferon-gamma (IFN-γ), a cytokine produced by sensitized T lymphocytes, is one of the key elements in defining T helper
1 lymphocyte immune responses. Quantitative evaluation of IFN-γ expression could provide an important analytical tool for
measurement of cell-mediated immunity and investigating immune responses to infectious diseases. Method of DNA-designed avian
IgY antibodies was used for production of monospecific polyclonal antibodies that allows quantification of the recombinant
bovine IFN-γ protein. IFN-γ cDNA was subcloned and expressed in mammalian expression plasmid (pcDNA3.1(+)) under the control
of the human cytomegalovirus promoter. Chickens were immunized by plasmid DNA, and eggyolk antibodies extracted from eggs
were collected after immunization. IgY-specific antibodies were evaluated by an antigen capture enzyme-linked immunosorbent
assay (ELISA) using recombinant IFN-γ. Based on the results, developed bovine IFN-γ capture ELISA could detect up to 1 ng/ml
of IFN-γ by 64-fold diluted IgY. Monospecific anti-bovine IFN-γ antibodies generated in chickens are useful for quantifying
different concentrations of recombinant bovine IFN-γ, which is expressed in cell culture. |
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