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Archives of Virology

, Volume 163, Issue 5, pp 1141–1152 | Cite as

Identification and production of mouse scFv to specific epitope of enterovirus-71 virion protein-2 (VP2)

  • Jeeraphong Thanongsaksrikul
  • Potjanee Srimanote
  • Pongsri Tongtawe
  • Kittirat Glab-ampai
  • Aijaz Ahmad Malik
  • Oratai Supasorn
  • Phatcharaporn Chiawwit
  • Yong Poovorawan
  • Wanpen Chaicumpa
Original Article

Abstract

Enterovirus-71 (EV71) and coxsackievirus-A16 (CA16) frequently cause hand-foot-mouth disease (HFMD) epidemics among infants and young children. CA16 infections are usually mild, while EV71 disease may be fatal due to neurologic complications. As such, the ability to rapidly and specifically recognize EV71 is needed to facilitate proper case management and epidemic control. Accordingly, the aim of this study was to generate antibodies to EV71-virion protein-2 (VP2) by phage display technology for further use in specific detection of EV71. A recombinant peptide sequence of EV71-VP2, carrying a predicted conserved B cell epitope fused to glutathione-S-transferase (GST) (designated GST-EV71-VP2/131-160), was produced. The fusion protein was used as bait in in-solution biopanning to separate protein-bound phages from a murine scFv (MuscFv) phage display library constructed from an immunoglobulin gene repertoire from naïve ICR mice. Three phage-transformed E. coli clones (clones 63, 82, and 83) produced MuscFvs that bound to the GST-EV71-VP2/131-160 peptide. The MuscFv of clone 83 (MuscFv83), which produced the highest ELISA signal to the target antigen, was further tested. MuscFv83 also bound to full-length EV71-VP2 and EV71 particles, but did not bind to GST, full-length EV71-VP1, or the antigenically related CA16. MuscFv83 could be a suitable reagent for rapid antigen-based immunoassay, such as immunochromatography (ICT), for the specific detection and/or diagnosis of EV71 infection as well as epidemic surveillance.

Notes

Acknowledgements

Funding was provided by grants from the following sources: Thailand Research Fund (MRG5680105); Thammasat University (Project IDs 218028, 180630, and 151632); Center of Excellence in Clinical Virology (GCE 58-014-30-004); Research Chair Grant (P-15-50004); and, NSTDA Chair Professor Grant (P-1450624). The authors gratefully acknowledge Ms Jiratchaya Puenpa of Chulalongkorn University for EV71 propagation.

Compliance with ethical standards

Consent for publication

All authors consent to publication.

Conflict of interest

All authors declare no personal or professional conflicts of interest, and no financial support from the companies that produce and/or distribute the drugs, devices, or materials described in this report.

Animal ethics

Institutional guidelines for the care and use of animals were followed.

Ethics approval and informed consent

This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

705_2018_3731_MOESM1_ESM.tif (10.1 mb)
Supplementary material 1 (TIFF 10314 kb) Supplementary Fig. 1 Computerized interaction of modeled-EV71-VP2 (genotype B5) and MuscFv63 with their interactive residues. (A) EV71-VP2 (grey) showing region predicted to contain the B-cell epitope (red). Sequence of the EV71 VP2 peptide is boxed; the EV71-VP2 specific residues are underlined; and the residues that were predicted to form contact interface with the MuscFv63 are bold. (B) MuscFv63 (green) protrudes VH-CDR loops to interact with the EV71-VP2 B-cell epitope (yellow). The VL did not interact with the target. (C) Interactive residues between MuscFv63 (green) and specific residues of EV71-VP2. H-bonds, grey dots
705_2018_3731_MOESM2_ESM.tif (10.9 mb)
Supplementary material 2 (TIFF 11135 kb) Supplementary Fig. S2 Computerized interaction between modeled-EV71-VP2 (genotype B5) and MuscFv82 with their interactive residues. (A) EV71-VP2 (grey) showing region predicted to contain the B-cell epitope (red) Sequence of the EV71 VP2 peptide is boxed; the EV71-VP2 specific residues are underlined, and the residues that were predicted to form contact interface with the MuscFv82 are bold. (B) As for the MuscFv63, the MuscFv82 used only CDRs of the VH domain to interact with the target. (C) Interactive residues between MuscFv82 (green) and specific residues of EV71-VP2. H-bonds, grey dots

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Jeeraphong Thanongsaksrikul
    • 1
  • Potjanee Srimanote
    • 1
  • Pongsri Tongtawe
    • 1
  • Kittirat Glab-ampai
    • 2
  • Aijaz Ahmad Malik
    • 3
  • Oratai Supasorn
    • 1
  • Phatcharaporn Chiawwit
    • 1
  • Yong Poovorawan
    • 4
  • Wanpen Chaicumpa
    • 2
  1. 1.Graduate Programme in Biomedical Science, Faculty of Allied Health SciencesThammasat UniversityRangsitThailand
  2. 2.Faculty of Medicine Siriraj HospitalCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Mahidol UniversityBangkokThailand
  3. 3.Faculty of Medical TechnologyCenter of Data Mining and Biomedical Informatics, Mahidol UniversityNakhon PathomThailand
  4. 4.Faculty of MedicineCenter of Excellence in Clinical Virology, Chulalongkorn UniversityBangkokThailand

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