Isolation and Characterization of scFv Antibody against Internal Ribosomal Entry Site of Enterovirus A71
Issued Date
2023-06-01
Resource Type
ISSN
16616596
eISSN
14220067
Scopus ID
2-s2.0-85163923781
Pubmed ID
37373012
Journal Title
International Journal of Molecular Sciences
Volume
24
Issue
12
Rights Holder(s)
SCOPUS
Bibliographic Citation
International Journal of Molecular Sciences Vol.24 No.12 (2023)
Suggested Citation
Hlaing S.T., Srimanote P., Tongtawe P., Khantisitthiporn O., Glab-ampai K., Chulanetra M., Thanongsaksrikul J. Isolation and Characterization of scFv Antibody against Internal Ribosomal Entry Site of Enterovirus A71. International Journal of Molecular Sciences Vol.24 No.12 (2023). doi:10.3390/ijms24129865 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/87866
Title
Isolation and Characterization of scFv Antibody against Internal Ribosomal Entry Site of Enterovirus A71
Author's Affiliation
Other Contributor(s)
Abstract
Enterovirus A71 (EV-A71) is one of the causative agents of hand-foot-mouth disease, which can be associated with neurocomplications of the central nervous system. A limited understanding of the virus’s biology and pathogenesis has led to the unavailability of effective anti-viral treatments. The EV-A71 RNA genome carries type I internal ribosomal entry site (IRES) at 5′ UTR that plays an essential role in the viral genomic translation. However, the detailed mechanism of IRES-mediated translation has not been elucidated. In this study, sequence analysis revealed that the domains IV, V, and VI of EV-A71 IRES contained the structurally conserved regions. The selected region was transcribed in vitro and labeled with biotin to use as an antigen for selecting the single-chain variable fragment (scFv) antibody from the naïve phage display library. The so-obtained scFv, namely, scFv #16-3, binds specifically to EV-A71 IRES. The molecular docking showed that the interaction between scFv #16-3 and EV-A71 IRES was mediated by the preferences of amino acid residues, including serine, tyrosine, glycine, lysine, and arginine on the antigen-binding sites contacted the nucleotides on the IRES domains IV and V. The so-produced scFv has the potential to develop as a structural biology tool to study the biology of the EV-A71 RNA genome.