Babesia Bovis: Detection and Characterization of Culture-Derived Exoantigens (Immunology, Parasitology)

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.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Veterinary Medical Science

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Montealegre, Federico Jose

• Biology, Veterinary Science

Identifier

(UMI)AAI8623373

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/71336