title: Characterization of periplasmic protein BP26 epitopes of Brucella melitensis reacting with murine monoclonal and sheep antibodies.
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.
authors.affiliation: Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
authors.forename: Jinlang
authors.initials: J
authors.lastname: Qiu
authors.forename: Wenjing
authors.initials: W
authors.lastname: Wang
authors.forename: Jingbo
authors.initials: J
authors.lastname: Wu
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Yuanzhi
authors.initials: Y
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Qiao
authors.forename: Chuangfu
authors.initials: C
authors.lastname: Chen
authors.forename: Goege F
authors.initials: GF
authors.lastname: Gao
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Allain
authors.forename: Chengyao
authors.initials: C
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D017347:Brucella melitensis / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal; C106040:BP26 protein, Brucella; D000939:Epitopes; D008565:Membrane Proteins