Babesia Bovis: Detection and Characterization of Culture-Derived Exoantigens (Immunology, Parasitology)
Montealegre, Federico Jose
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https://hdl.handle.net/2142/71336
Description
Title
Babesia Bovis: Detection and Characterization of Culture-Derived Exoantigens (Immunology, Parasitology)
Author(s)
Montealegre, Federico Jose
Issue Date
1986
Department of Study
Veterinary Medical Science
Discipline
Veterinary Medical Science
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Veterinary Science
Abstract
A two-site enzyme-immunoassay was developed to detect culture-derived Babesia bovis exoantigens. Bovine anti-B. bovis IgG was used as a capture antibody and as a recognizing antibody. A capture antibody concentration of 10 ug/ml and test samples at protein concentrations of 100 ug/ml gave optimal results with the horseradish peroxidase-conjugated bovine anti-B. bovis IgG. The test was sensitive, specific and reproducible. The highest antigenic activity was demonstrated in 24-hour continuous B. bovis cultures in the microaerophilous stationary phase (MASP) system. The test also proved capable of detecting cross-reactivity with Babesia bigemina. Anion-exchange chromatography with a pH gradient was used as a first-step fractionation of B. bovis exoantigens. Most of the antigenic activity was detected at pH 4. Subsequent re-chromatography of this fraction was conducted using size exclusion high performance liquid chromatography (HPLC). These antigens were detected in the 200 Kd peak. Analysis of the partially purified pH 4 using enzyme-linked immunotransfer blot demonstrated electronegative antigens of relative molecular weights of 57 Kd, 55 Kd and 43 K respectively. The major antigenic bands were confirmed in the partially purified HPLC fraction. A purity analysis of the partially purified fractions showed at least two major contaminant groups (gamma and beta proteins) were present.
The potential applications of the techniques of detection and isolation are discussed in relation to diagnosis and immunoprophylaxis of bovine babesiosis.
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