1. Identification of two novel B cell epitopes on E184L protein of African swine fever virus using monoclonal antibodies.
- Author
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Tesfagaber W, Lan D, Wang W, Zhao R, Yin L, Yang M, Zhu Y, Sun E, Liu R, Lin W, Bu Z, Li F, and Zhao D
- Subjects
- Animals, Swine, African Swine Fever immunology, African Swine Fever virology, Mice, Viral Proteins immunology, Viral Proteins genetics, Viral Proteins chemistry, Antigens, Viral immunology, Antigens, Viral genetics, Antigens, Viral chemistry, Mice, Inbred BALB C, African Swine Fever Virus immunology, African Swine Fever Virus genetics, Antibodies, Monoclonal immunology, Epitopes, B-Lymphocyte immunology, Epitope Mapping, Antibodies, Viral immunology
- Abstract
African swine fever virus (ASFV) is a large double-stranded DNA virus with a complex structural architecture and encodes more than 150 proteins, where many are with unknown functions. E184L has been reported as one of the immunogenic ASFV proteins that may contribute to ASFV pathogenesis and immune evasion. However, the antigenic epitopes of E184L are not yet characterized. In this study, recombinant E184L protein was expressed in prokaryotic expression system and four monoclonal antibodies (mAbs), designated as 1A10, 2D2, 3H6, and 4C10 were generated. All four mAbs reacted specifically with ASFV infected cells. To identify the epitopes of the mAbs, a series of overlapped peptides of E184L were designed and expressed as maltose binding fusion proteins. Accordingly, the expressed fusion proteins were probed with each E184L mAb separately by using Western blot. Following a fine mapping, the minimal linear epitope recognized by mAb 1A10 was identified as
119 IQRQGFL125 , and mAbs 2D2, 3H6, and 4C10 recognized a region located between153 DPTEFF158 . Alignment of amino acids of E184L revealed that the two linear epitopes are highly conserved among different ASFV isolates. Furthermore, the potential application of the two epitopes in ASFV diagnosis was assessed through epitope-based ELISA using 24 ASFV positive and 18 negative pig serum and the method were able to distinguish positive and negative samples, indicating the two epitopes are dominant antigenic sites. To our knowledge, this is the first study to characterize the B cell epitopes of the antigenic E184L protein of ASFV, offering valuable tools for future research, as well as laying a foundation for serological diagnosis and epitope-based marker vaccine development., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)- Published
- 2024
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