In vitro characterization of neutralizing hen antibodies to coxsackievirus a16

Pharaoh Fellow Mwale, Chi Hsin Lee, Peng Nien Huang, Sung Nien Tseng, Shin Ru Shih, Hsin Yuan Huang, Sy Jye Leu, Yun Ju Huang, Liao Chun Chiang, Yan Chiao Mao, Wei Chu Wang, Yi Yuan Yang

Research output: Contribution to journalArticlepeer-review

Abstract

Coxsackievirus A16 (CA16) is one of the major causative agents of hand, foot, and mouth disease (HFMD). Children aged <5 years are the most affected by CA16 HFMD globally. Although clinical symptoms of CA16 infections are usually mild, severe complications, such as aseptic meningitis or even death, have been recorded. Currently, no vaccine or antiviral therapy for CA16 infection exists. Single‐chain variable fragment (scFv) antibodies significantly inhibit viral infection and could be a potential treatment for controlling the infection. In this study, scFv phage display libraries were constructed from splenocytes of a laying hen immunized with CA16‐infected lysate. The pComb3X vector containing the scFv genes was introduced into ER2738 Escherichia coli and rescued by helper phages to express scFv molecules. After screening with five cycles of bio‐panning, an effective scFv antibody showing favorable binding activity to proteins in CA16‐infected lysate on ELISA plates was selected. Importantly, the selected scFv clone showed a neutralizing capability against the CA16 virus and cross‐reacted with viral proteins in EV71‐infected lysate. Intriguingly, polyclonal IgY antibody not only showed binding specificity against proteins in CA16‐infected lysate but also showed significant neutralization activities. Nevertheless, IgY‐binding protein did not cross‐react with proteins in EV71‐infected lysate. These results suggest that the IgY‐ and scFv-binding protein antibodies provide protection against CA16 viral infection in in vitro assays and may be potential candidates for treating CA16 infection in vulnerable young children.

Original languageEnglish
Article number4146
JournalInternational journal of molecular sciences
Volume22
Issue number8
DOIs
Publication statusPublished - Apr 2 2021

Keywords

  • Coxsackievirus A16 (CA16)
  • Enterovirus 71 (EV71)
  • Immunoglobulin yolk (IgY)
  • Phage display libraries
  • Single‐chain variable fragment (scFv)

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

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