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Published on: March 24, 2017
Monoclonal Antibody Targeting a Conserved N-Terminal Epitope on 2Apro of Enteroviruses
Kai Yi1,2, Lilan Xie1,2, Yaoming Li1,2
1Hubei Engineering Research Center of Viral Vector, Applied Biotechnology Research Center, Wuhan University of Bioengineering, Wuhan, China.
Abstract:
Several members of enteroviruses (EVs) that belong to the EVs A and B species cause hand, foot, and mouth disease (HFMD) in infants and young children. The virus-specific protease 2Apro is conserved in all the EV species, thus developing a monoclonal antibody (mAb) against 2Apro may facilitate the identification from the HFMD-associated pathogens. In this study, we achieved a murine mAb, named 5A3, specifically toward EVA71 2Apro by using the traditional hybridoma technique. The mAb 5A3 recognizes 2Apro of both EVs A and B species, which was demonstrated by indirect fluorescent assay and Western blotting. Furthermore, a conserved N-terminal epitope on 2Apro recognized by mAb 5A3 was defined by using an overlapping peptide-based enzyme-linked immunosorbent assay (ELISA). Therefore, the unique mAb targeting conserved EVs 2Apro can be used as an important tool during both identifying the causative agent of HFMD and elucidating the pathological mechanism of HFMD.
Insights
Researchers developed a new monoclonal antibody (mAb) targeting enterovirus (EV) protease 2Apro. This antibody aids in identifying hand, foot, and mouth disease pathogens and understanding disease mechanisms.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Enteroviruses (EVs) from species A and B are common causes of hand, foot, and mouth disease (HFMD) in children.
- The viral protease 2Apro is a conserved protein across EV species, making it a potential target for diagnostics.
Purpose of the Study:
- To develop a monoclonal antibody (mAb) against the conserved enterovirus protease 2Apro.
- To evaluate the utility of this mAb in identifying HFMD-associated pathogens.
Main Methods:
- Hybridoma technique was used to generate a murine mAb, designated 5A3.
- Indirect fluorescent assay and Western blotting were employed to assess mAb reactivity.
- Overlapping peptide-based ELISA was utilized to define the epitope recognized by the mAb.
Main Results:
- The mAb 5A3 was successfully generated and specifically targets EVA71 2Apro.
- mAb 5A3 demonstrated cross-reactivity with 2Apro from both Enterovirus species A and B.
- A conserved N-terminal epitope on 2Apro was identified as the target of mAb 5A3.
Conclusions:
- The developed mAb 5A3 is a valuable tool for identifying enteroviruses causing HFMD.
- This mAb can aid in elucidating the pathological mechanisms of HFMD.
- Targeting conserved viral proteins like 2Apro offers a promising strategy for diagnostic development.
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