Monoclonal antibody against Babesia equi: characterization and potential application of antigen for serodiagnosis.
Abstract: Monoclonal antibody (MAb) BEG3 was produced against Babesia equi parasites to define a species-specific antigen for diagnostic use. The MAb reacted with single, paired, and Maltese cross forms of B. equi, and no reaction was observed with this MAb on acetone-fixed Babesia caballi, Babesia ovata, or Babesia microti parasites in the indirect immunofluorescent antibody test. Confocal laser and immunoelectron microscopic studies showed that the antigen which was recognized by this MAb was located on the surface of B. equi parasites. This MAb recognized a 19-kDa protein of B. equi antigen and did not react with B. caballi antigen or normal horse erythrocytes in immunoblot analysis. This MAb also significantly inhibited the in vitro growth of the B. equi parasite. Preliminary studies using partially purified antigen, which was separated by high-pressure liquid chromatography and recognized by the MAb, suggested that it is a suitable antigen for enzyme-linked immunosorbent assay detection of anti-B. equi antibodies in naturally infected horse sera.
Publication Date: 1998-07-03 PubMed ID: 9650921PubMed Central: PMC104937DOI: 10.1128/JCM.36.7.1835-1839.1998Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Antigen
- Diagnosis
- Disease Diagnosis
- Enzyme-Linked Immunosorbent Assay (ELISA)
- Equine Health
- Erythrocytes
- High-performance Liquid Chromatography (HPLC)
- Immunoblotting
- Immunofluorescence Assay
- Immunohistochemistry
- Immunology
- In Vitro Research
- Infectious Disease
- Microscopy
- Monoclonal Antibodies
- Parasites
- Protein
- Serodiagnosis
- Theileria equi
- Veterinary Medicine
- Veterinary Research
Summary
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The researchers present a study on the production of Monoclonal antibody, named BEG3, which specifically targets a particular species of parasites, Babesia equi. Using this antibody, the investigators have identified a potentially useful tool that can be used to diagnose the presence of these parasites in horses.
Monoclonal Antibody Production
- The study revolves around the production of a specific Monoclonal antibody (MAb) known as BEG3, created to react against Babesia equi – a parasite that infects red blood cells in horses leading to a disease condition called Equine Piroplasmosis.
- The manufacturing of this MAb helps to identify a unique, species-specific antigen that could be useful for diagnostic purposes.
Reaction with Various Forms of B. equi
- Upon trials, this MAb showed a positive response with all forms of B. equi – single, paired, and a particular arrangement known as the Maltese cross.
- Interestingly, no reaction was seen when this MAb was exposed to other related parasite species including Babesia caballi, Babesia ovata, or Babesia microti in the indirect immunofluorescent antibody test – indicating BEG3’s selectivity for B. equi.
Location of Antigen Recognition
- The study further employed microscopy techniques such as confocal laser and immunoelectron microscopy which revealed that the antigen to which the MAb reacts is located on the surface of the B. equi parasites.
Protein Recognition and Restriction
- Immunoblot analysis showed that the MAb recognizes a 19 kiloDalton (kDa) protein associated with B. equi.
- Interestingly, BEG3 did not react with either antigens from B. caballi or with normal horse red blood cells.
- Furthermore, it was observed that the BEG3 MAb could significantly inhibit the in vitro (laboratory conditions) growth of B. equi parasites, marking its potential use in therapeutic interventions.
Application in Diagnostic Testing
- Finally, preliminary studies were carried out that used a partially purified antigen (separated via high-pressure liquid chromatography) which was recognizable by the BEG3 MAb.
- The results from this part of the study suggest the antigen’s suitability in developing enzyme-linked immunosorbent assays (ELISAs) for detecting the presence of anti-B. equi antibodies in horse sera, serving as a diagnostic tool.
Cite This Article
APA
Avarzed A, Igarashi I, De Waal DT, Kawai S, Oomori Y, Inoue N, Maki Y, Omata Y, Saito A, Nagasawa H, Toyoda Y, Suzuki N.
(1998).
Monoclonal antibody against Babesia equi: characterization and potential application of antigen for serodiagnosis.
J Clin Microbiol, 36(7), 1835-1839.
https://doi.org/10.1128/JCM.36.7.1835-1839.1998 Publication
Researcher Affiliations
- The Research Center for Protozoan Molecular Immunology, Obihiro University of Agriculture and Veterinary Medicine, Hokkaido, Japan.
MeSH Terms
- Animals
- Antibodies, Monoclonal / biosynthesis
- Antibodies, Monoclonal / immunology
- Antibodies, Monoclonal / isolation & purification
- Antibodies, Protozoan / blood
- Antigens, Protozoan / analysis
- Antigens, Protozoan / immunology
- Antigens, Protozoan / isolation & purification
- Babesia / growth & development
- Babesia / immunology
- Babesia / isolation & purification
- Babesiosis / diagnosis
- Babesiosis / parasitology
- Electrophoresis, Polyacrylamide Gel
- Enzyme-Linked Immunosorbent Assay
- Erythrocytes / parasitology
- Fluorescent Antibody Technique, Indirect
- Horse Diseases / diagnosis
- Horse Diseases / parasitology
- Horses
- Immune Sera
- Immunoblotting
- Microscopy, Confocal
- Microscopy, Immunoelectron
- Species Specificity
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Citations
This article has been cited 7 times.- Tuvshintulga B, Nugraha AB, Mizutani T, Liu M, Ishizaki T, Sivakumar T, Xuan X, Yokoyama N, Igarashi I. Development of a stable transgenic Theileria equi parasite expressing an enhanced green fluorescent protein/blasticidin S deaminase.. Sci Rep 2021 Apr 27;11(1):9107.
- El-Shehabi F, Vermeire JJ, Yoshino TP, Ribeiro P. Developmental expression analysis and immunolocalization of a biogenic amine receptor in Schistosoma mansoni.. Exp Parasitol 2009 May;122(1):17-27.
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- Yokoyama N, Bork S, Nishisaka M, Hirata H, Matsuo T, Inoue N, Xuan X, Suzuki H, Sugimoto C, Igarashi I. Roles of the Maltese cross form in the development of parasitemia and protection against Babesia microti infection in mice.. Infect Immun 2003 Jan;71(1):411-7.
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