Characterization of a murine monoclonal antibody generated against Neospora caninum tachyzoites by use of western blot analysis and immunoelectron microscopy

Rebecca A. Cole From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849 (Cole, Lindsay, Toivio-Kinnucan, Blagburn), and USDA, Agriculture Research Service, Livestock and Poultry Science Institute Zoonotic Diseases Laboratory, Beltsville, MD 20705 (Dubey).

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David S. Lindsay From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849 (Cole, Lindsay, Toivio-Kinnucan, Blagburn), and USDA, Agriculture Research Service, Livestock and Poultry Science Institute Zoonotic Diseases Laboratory, Beltsville, MD 20705 (Dubey).

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J. P. Dubey From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849 (Cole, Lindsay, Toivio-Kinnucan, Blagburn), and USDA, Agriculture Research Service, Livestock and Poultry Science Institute Zoonotic Diseases Laboratory, Beltsville, MD 20705 (Dubey).

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Maria A. Toivio-Kinnucan From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849 (Cole, Lindsay, Toivio-Kinnucan, Blagburn), and USDA, Agriculture Research Service, Livestock and Poultry Science Institute Zoonotic Diseases Laboratory, Beltsville, MD 20705 (Dubey).

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Byron L. Blagburn From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849 (Cole, Lindsay, Toivio-Kinnucan, Blagburn), and USDA, Agriculture Research Service, Livestock and Poultry Science Institute Zoonotic Diseases Laboratory, Beltsville, MD 20705 (Dubey).

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Summary

A murine monoclonal antibody (mab) 6G7 generated against tachyzoites of Neospora caninum recognized 8 major and several minor antigens, as observed by western blot analysis. Relative rate of migration of the 8 major antigens ranged from 31 to 97.4 kd. In addition, mab 6G7 recognized a Toxoplasma gondii tachyzoite antigen with a relative rate of migration of 107 kd. Immunogold labeling of N caninum tachyzoites grown in human foreskin fibroblast cells indicated that mab 6G7 binds to micronemes, dense granules, basal portions of rhoptries, and intravacuolar tubules within the parasitophorous vacuole. Monoclonal antibody 6G7 also bound to micronemes and basal portions of rhoptries within tachyzoites of T gondii. Monoclonal antibody 6G7 did not significantly inhibit development of tachyzoites in vitro.

Summary

A murine monoclonal antibody (mab) 6G7 generated against tachyzoites of Neospora caninum recognized 8 major and several minor antigens, as observed by western blot analysis. Relative rate of migration of the 8 major antigens ranged from 31 to 97.4 kd. In addition, mab 6G7 recognized a Toxoplasma gondii tachyzoite antigen with a relative rate of migration of 107 kd. Immunogold labeling of N caninum tachyzoites grown in human foreskin fibroblast cells indicated that mab 6G7 binds to micronemes, dense granules, basal portions of rhoptries, and intravacuolar tubules within the parasitophorous vacuole. Monoclonal antibody 6G7 also bound to micronemes and basal portions of rhoptries within tachyzoites of T gondii. Monoclonal antibody 6G7 did not significantly inhibit development of tachyzoites in vitro.

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