利用单克隆抗体对羊种布鲁氏菌新型Omp31抗原表位进行鉴定

Characterization of novel Omp31 antigenic epitopes of Brucella melitensis by monoclonal antibodies.

作者信息

Li Jinfeng, Hu Feihuan, Chen Shouyi, Luo Peifang, He Zuoping, Wang Wenjing, Allain Jean-Pierre, Li Chengyao

机构信息

Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.

Guangzhou Center of Disease Control and Prevention (CDC), Guangzhou, China.

出版信息

BMC Microbiol. 2017 May 15;17(1):115. doi: 10.1186/s12866-017-1025-3.

BACKGROUND

Brucellosis is a severe zoonotic disease worldwide. Detection and identification of Brucella species are essential to prevent or treat brucellosis in humans and animals. The outer membrane protein-31 (Omp31) is a major protein of Brucellae except for B. abortus, while the Omp31 antigenic epitopes have not been extensively characterized yet.

RESULTS

A total of 22 monoclonal antibodies (mAbs) were produced against Omp31 of Brucella (B.) melitensis, of which 13 recognized five linear epitopes, 7 reacted with semi-conformational epitopes and 2 reacted with conformational epitopes, respectively. The mAb isotypes were 11 (50%) IgG2a, 5 (23%) IgG1 and 6 (27%) IgM. On the basis of epitope recognition and reactivity levels, 8 mAbs including 3 IgM and 5 IgG clones were considered as highly reactive and potentially diagnostic antibodies. Among these mAbs, 7A3 (IgG1), 5B1 (IgG2a), 2C1 (IgG2a) and 5B3 (IgG2a) reacted with differently conserved linear epitopes of B. melitensis, B. ovis, B. suis and B. canis strains, while 5H3 (IgG2a) highly reacted with a conformational epitope of Omp31 when tested with several immunoassays.

CONCLUSIONS

These potent monoclonal antibodies can be used for identifying Omp31 antigens or detecting B. melitensis and other Brucella species beyond B. abortus in vitro or in vivo.

背景

布鲁氏菌病是一种在全球范围内严重的人畜共患病。布鲁氏菌属物种的检测和鉴定对于预防或治疗人和动物的布鲁氏菌病至关重要。外膜蛋白-31（Omp31）是除流产布鲁氏菌外布鲁氏菌的主要蛋白，而Omp31抗原表位尚未得到广泛表征。

结果

共产生了22种针对羊种布鲁氏菌Omp31的单克隆抗体（mAb），其中13种识别5个线性表位，7种与半构象表位反应，2种与构象表位反应。mAb的亚型为11种（50%）IgG2a、5种（23%）IgG1和6种（27%）IgM。基于表位识别和反应水平，8种mAb（包括3种IgM和5种IgG克隆）被认为是高反应性和潜在诊断性抗体。在这些mAb中，7A3（IgG1）、5B1（IgG2a）、2C1（IgG2a）和5B3（IgG2a）与羊种布鲁氏菌、绵羊布鲁氏菌、猪种布鲁氏菌和犬种布鲁氏菌菌株不同保守的线性表位反应，而5H3（IgG2a）在用几种免疫测定法检测时与Omp31的构象表位高度反应。