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Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
Characterization of novel Omp31 antigenic epitopes of Brucella melitensis by monoclonal antibodies
Jinfeng Li1, Feihuan Hu1, Shouyi Chen2
1Department of Transfusion Medicine, Southern Medical University, Guangzhou, China.
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.
Insights
Researchers developed potent monoclonal antibodies targeting the Brucella outer membrane protein-31 (Omp31). These antibodies are crucial for accurately detecting and identifying Brucella species, aiding in the diagnosis of this severe zoonotic disease.
Area of Science:
- Immunology
- Microbiology
- Veterinary Science
Background:
- Brucellosis is a significant global zoonotic disease affecting both humans and animals.
- Accurate detection and identification of Brucella species are critical for disease management.
- The outer membrane protein-31 (Omp31) is a key antigen in Brucella, but its epitopes require further characterization.
Purpose of the Study:
- To produce and characterize monoclonal antibodies (mAbs) against the Omp31 protein of Brucella melitensis.
- To identify specific linear, semi-conformational, and conformational epitopes on Omp31.
- To evaluate the diagnostic potential of generated mAbs for Brucella detection.
Main Methods:
- Production of 22 monoclonal antibodies against Brucella melitensis Omp31.
- Epitope mapping using various immunoassays to determine antibody binding characteristics.
- Analysis of antibody isotypes (IgG2a, IgG1, IgM) and reactivity levels.
- Testing antibody cross-reactivity with different Brucella species.
Main Results:
- 13 mAbs recognized linear epitopes, 7 reacted with semi-conformational epitopes, and 2 recognized conformational epitopes.
- Eight highly reactive mAbs (3 IgM, 5 IgG) were identified as potentially diagnostic.
- Specific mAbs demonstrated reactivity with conserved epitopes across multiple Brucella species, including B. melitensis, B. ovis, B. suis, and B. canis.
- One mAb (5H3) showed high reactivity with a conformational epitope on Omp31.
Conclusions:
- Developed potent monoclonal antibodies targeting Brucella Omp31.
- These antibodies can be utilized for the identification of Omp31 antigens.
- The mAbs enable the detection of Brucella melitensis and other Brucella species (excluding B. abortus) in various settings.
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