Document Type : Short Communication

Authors

1 Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

2 Department of Pathobiology, Faculty of Veterinary Medicine, University of Shahrekord, Shahrekord, Iran

3 DVM Graduate, School of Veterinary Medicine, Shiraz University, Shiraz, Iran

4 Department of Biotechnology, GLA University, Mathura, Uttar Pradesh, India

Abstract

No diagnostic kits and reagents are available in the market to detect and evaluate camel immune responses to different pathogens. This study aimed to produce sheep anti-camel (Camelus dromedarius) polyclonal antibodies (pAbs) and to determine the specificity with other species immunoglobulin. Immunoglobulins (Igs) from camel serum samples were purified using ammonium sulfate precipitation (40.00% saturated ammonium sulfate). Purity of the camel Igs was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis. PAbs against (Camelus dromedarius) immunoglobulins were generated by immunizing sheep with purified Igs. Anti- camel Ig polyclonal antibodies titer and specificity were determined using ELISA and Western blot techniques. Polyclonal antibodies specific to camel Igs were significantly high in immunized sheep which confirmed the immunization procedure. PAbs reacted specifically with camel serum immunoglobulin and did not react with other species immunoglobulin of horse and chickens. Polyclonal antibodies produced in this study can be regarded as a valuable tool to be used for immune-diagnostic purposes in camel population world-wide.

Keywords

  1. Padlan Anatomy of the antibody molecule. Mol Immunol 1994; 31(3): 169-217.
  2. Azwai SM, Carter SD, Woldehiwet Z. The isolation and characterization of camel (Camelus dromedarius) immunoglobulin classes and subclasses. J Comp Pathol 1993; 109(2): 187-195.
  3. Azwai SM, Carter SD Woldehiwet Z. Monoclonal antibodies against camel (Camelus dromedarius) IgG, IgM and light chains. Vet Immunol Immunopathol 1995; 45(1-2): 175-184.
  4. Azwai SM, Carter SD, Woldehiwet Z. Immunoglobulins of Camel (Camelus dromedarius) Colostrum. J Comp Pathol 1996; 114(3): 273-282.
  5. Hamers-Casterman C, Atarhouch T, Muyldermans S, et al. Naturally occurring antibodies devoid of light chains. Nature 1993; 363(6428): 446-448.
  6. Heddle RJ, Rowley D. Dog immunoglobulins. I. immunochemical characterization of dog serum, parotid saliva, colostrum, milk and small bowel fluid. Immunology 1975; 29(1): 185-195.
  7. Laemmli Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259): 680-685.
  8. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72: 248-254.
  9. Harlow E, Lane D. Antibodies: A Laboratory manual. New York, USA: Cold spring harbor laboratory press 1988; 680-685.
  10. Aghebati-Maleki L, Bakhshinejad B, Baradaran B, et al. Phage display as a promising approach for vaccine development. J Biomed Sci 2016; 23(1): 66. doi: 10.1186/s12929-016-0285-9.
  11. Leusen JH, Nimmerjahn F. The role of IgG in immune responses. In: Falk N (Ed). Molecular and cellular mechanisms of antibody activity. New York, USA: Springer 2013; 85-112.
  12. Kinyanjui SM, Mwangi T, Bull PC, et al. Protection against clinical malaria by heterologous immunoglobulin G antibodies against malaria-infected erythrocyte variant surface antigens requires interaction with asymptomatic infections. J Infect Dis 2004; 190(9): 1527-1533.
  13. Stanisic DI, Richards JS, McCallum FJ, et al. Immunoglobulin G subclass-specific responses against Plasmodium falciparum merozoite antigens are associated with control of parasitemia and protection from symptomatic illness. Infect Immun 2009; 77(3):1165-1174.
  14. Abdel-Rahman EH, El-Jakee JK, Hatem ME, et al. Preparation of goat and rabbit anti-camel immuno-globulin G whole molecule labeled with horseradish peroxidase. Vet World 2017; 10(1): 92-100.
  15. Ezzatifar F, Majidi J, Baradaran B, et al. Development and characterization of monoclonal antibodies against human IgA in Balb/c mice. Hum Antibodies 2014; 23(1-2): 7-12.