Characterization of Periplasmic Protein BP26 Epitopes of Brucella melitensis Reacting with Murine Monoclonal and Sheep Antibodies
Open Access
- 23 March 2012
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 7 (3), e34246
- https://doi.org/10.1371/journal.pone.0034246
Abstract
More than 35,000 new cases of human brucellosis were reported in 2010 by the Chinese Center for Disease Control and Prevention. An attenuated B. melitensis vaccine M5-90 is currently used for vaccination of sheep and goats in China. In the study, a periplasmic protein BP26 from M5-90 was characterized for its epitope reactivity with mouse monoclonal and sheep antibodies. A total of 29 monoclonal antibodies (mAbs) against recombinant BP26 (rBP26) were produced, which were tested for reactivity with a panel of BP26 peptides, three truncated rBP26 and native BP26 containing membrane protein extracts (NMP) of B. melitensis M5-90 in ELISA and Western-Blot. The linear, semi-conformational and conformational epitopes from native BP26 were identified. Two linear epitopes recognized by mAbs were revealed by 28 of 16mer overlapping peptides, which were accurately mapped as the core motif of amino acid residues 93DRDLQTGGI101 (position 93 to 101) or residues 104QPIYVYPD111, respectively. The reactivity of linear epitope peptides, rBP26 and NMP was tested with 137 sheep sera by ELISAs, of which the two linear epitopes had 65–70% reactivity and NMP 90% consistent with the results of a combination of two standard serological tests. The results were helpful for evaluating the reactivity of BP26 antigen in M5-90.This publication has 29 references indexed in Scilit:
- Expression, purification, and improved antigenic specificity of a truncated recombinant bp26 protein of Brucella melitensis M5‐90: a potential antigen for differential serodiagnosis of brucellosis in sheep and goatsBiotechnology and Applied Biochemistry, 2011
- Human Brucellosis, Inner Mongolia, ChinaEmerging Infectious Diseases, 2010
- Association of anti-E1E2 antibodies with spontaneous recovery or sustained viral response to therapy in patients infected with hepatitis C virusJournal of Hepatology, 2010
- Brucellosis: The case for live, attenuated vaccinesVaccine, 2009
- Immunization with Recombinant Brucella Species Outer Membrane Protein Omp16 or Omp19 in Adjuvant Induces Specific CD4 + and CD8 + T Cells as Well as Systemic and Oral Protection against Brucella abortus InfectionInfection and Immunity, 2009
- Selection of Protective Epitopes for Brucella melitensis by DNA VaccinationInfection and Immunity, 2005
- Epitope Mapping of the Brucella melitensis BP26 Immunogenic Protein: Usefulness for Diagnosis of Sheep BrucellosisClinical and Vaccine Immunology, 2003
- Production and Characterization of Monoclonal Antibodies Specific for a Conserved Epitope within Hepatitis C Virus Hypervariable Region 1Journal of Virology, 2001
- Use of Recombinant BP26 Protein in Serological Diagnosis of Brucella melitensis Infection in SheepClinical and Diagnostic Laboratory Immunology, 2001
- Cloning, nucleotide sequence, and expression of the Brucella melitensis bp26 gene coding for a protein immunogenic in infected sheepFEMS Microbiology Letters, 1996