Production of a monoclonal antibody against chicken immunoglobulin G: A valuable molecule with research and diagnostic applications

Document Type: Original Article

Authors

1 Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

2 Department of Immunology, School of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

3 Department of Pathobiology, Faculty of Veterinary Science, Bu-Ali Sina University, Hamedan, Iran

4 Graduate student, Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, , Ahvaz, Iran

Abstract

Monoclonal antibodies (MAbs) are invaluable molecules which have several advantages over polyclonal immunoglobulins (Igs) including consistency and higher specificity and hence can be used in biological researches, diagnosis and treatment of diseases. The present study was conducted to produce monoclonal antibody against chicken IgG.TheIgG molecules were purified from chicken serum and used as antigens to immunize several mice. Thereafter, a well-immunized mouse was chosen and used for fusion process. After production of hybridoma cells, several rounds of cloning were carried out and produced MAbs were examined by various immunological assays including enzyme-linked immunosorbent assay (ELISA) and western and dot blotting. Assessment of grown hybridomas indicated that only one clone (5B8) has produced desired MAb against chicken IgG. Meanwhile, using an indirect ELISA, it was shown that this MAb successfully recognizes chicken IgG molecules attached to influenza virus nucleoprotein. Evaluation of cross reactivity of MAb 5B8 with several avian serum samples revealed that this molecule specifically identifies chicken antibody molecules. However, it also recognized turkey antibodies with less affinity. In addition to research applications like isolation and purification of chicken and turkey IgG molecules, such a MAb can be applied to design and development of various immunoassays (e.g. ELISA) in these avian species.

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  1. Warr GW, Magor KE, Higgins DA. IgY: Clues to the origins of modern antibodies. ImmunolToday 1995; 16(8): 392-398.
  2. Myers TJ, Schat KA, Mockett AP. Development of immunoglobulin class-specific enzyme-linked immunosorbent assays for measuring antibodies against avian rotavirus. Avian Dis 1989; 33(1): 53-59.
  3. Leenaars M, Hendriksen CF. Critical steps in the production of polyclonal and monoclonal antibodies: evaluation and recommendations. ILAR J 2005; 46(3): 269-279.
  4. Lipman NS, Jackson LR, Trudel LJ, et al. Monoclonal versus polyclonal antibodies: Distinguishing characteristics, applications, and information resources. ILAR J 2005; 46(3): 258-268.
  5. Pandey S. Hybridoma technology for production of monoclonal antibodies. Int J Pharm Sci Rev Res 2010; 1(2): 88-94.
  6. Schimizu M, Watanabe A, Tanaka A. Detection of high molecular weight mucin-like glycoprotein-A (HMGP-A) of human milk by chicken egg yolk antibody. Bioscien Biotechnol Biochem 1995; 59(1): 138-142.
  7. Wooley JA, Landon J. Comparison of antibody production to human interleukin-6 (IL-6) by sheep and chickens. J Immunol Methods 1995; 178(2): 253-265.
  8. Hodek P, Stiborova M. Chicken antibodies superior alternative for conventional immunoglobulins. Proc Indian Natl Sci Acad 2003; 69(4): 461-468.
  9. Raj GD, Latha B, Chandrasekhar MS, et al. Production, characterization and application of monoclonal antibodies against chicken IgY. Vet Arch 2004; 74(3): 189-199.
  10. Narat M, Bicek A, Vadnjal R, et al. Production, characterization and use of monoclonal antibodies recognizing IgY epitopes shared by chicken, turkey, pheasant, peafowl and sparrow. Food Technol Biotechnol 2004; 42(3): 175-182.
  11. Bhanushali JK, Gilbert JM, McDougald LR. Simple method to purify chicken immunoglobulin G. Poult Sci 1994; 73(7): 1158-1161.
  12. Harlow E, Lane D. Antibodies: A Laboratory Manual. New York, USA: Cold Spring Harbor Laboratory Press 1988; 139-245.
  13. Ausubel FM, Brent R, Kingston RE, et al. Short protocols in molecular biology. 2nd ed. New York, USA: John Willey and Sons 1995; 130-135.
  14. Jaydari A. Seyfi Abad Shapouri MR. Development of enzyme-linked immunosorbent assay with a nucleoprotein as an antigen for detection of antibodies against avian influenza virus. In proceedings: 15th International Congress of Microbiology. Tehran, Iran. 2014; 72.
  15. Erhard MH, Von Quistrop I, Junglling A, et al. Development of specific enzyme-linked immuno-sorbent antibody assay systems for the detection of chicken immunoglobulins G, M, and A using monoclonal antibodies. Poult Sci 1992; 71(2): 302-310.