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Development of DNA-Designed Avian IgY Antibodies for Quantitative Determination of Bovine Interferon-Gamma

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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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Authors and Affiliations

  1. Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Gholamreza Nikbakht Brujeni

  2. Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Sayed Amir Hossein Jalali

  3. Department of Toxicology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

    Mohammad Kazem Koohi

Authors
  1. Gholamreza Nikbakht Brujeni
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  2. Sayed Amir Hossein Jalali
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  3. Mohammad Kazem Koohi
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Correspondence to Gholamreza Nikbakht Brujeni.

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Nikbakht Brujeni, G., Jalali, S.A.H. & Koohi, M.K. Development of DNA-Designed Avian IgY Antibodies for Quantitative Determination of Bovine Interferon-Gamma. Appl Biochem Biotechnol 163, 338–345 (2011). https://doi.org/10.1007/s12010-010-9042-9

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  • DOI: https://doi.org/10.1007/s12010-010-9042-9

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