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Article

In Vitro Characterization of Neutralizing Hen Antibodies to Coxsackievirus A16

1
Ph.D. Program in Medical Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan
2
School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 110301, Taiwan
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Division of Infectious Diseases, Department of Pediatrics, Linkou Chang Gung Memorial Hospital, Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan 333423, Taiwan
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Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan 333323, Taiwan
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Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan 333423, Taiwan
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Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei 110301, Taiwan
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Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 110301, Taiwan
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Institute of Bioinformatics and Structural Biology, College of Life Sciences, National Tsing Hua University, Hsinchu 300040, Taiwan
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Division of Clinical Toxicology, Department of Emergency Medicine, Taichung Veterans General Hospital, Taichung 407219, Taiwan
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Core Laboratory of Antibody Generation and Research, Taipei Medical University, Taipei 110301, Taiwan
*
Author to whom correspondence should be addressed.
Academic Editor: Marc H.V. Van Regenmortel
Int. J. Mol. Sci. 2021, 22(8), 4146; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084146
Received: 12 March 2021 / Revised: 14 April 2021 / Accepted: 14 April 2021 / Published: 16 April 2021
(This article belongs to the Collection Feature Papers in Molecular Microbiology)
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. View Full-Text
Keywords: Coxsackievirus A16 (CA16); single-chain variable fragment (scFv); phage display libraries; Enterovirus 71 (EV71); immunoglobulin yolk (IgY) Coxsackievirus A16 (CA16); single-chain variable fragment (scFv); phage display libraries; Enterovirus 71 (EV71); immunoglobulin yolk (IgY)
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MDPI and ACS Style

Mwale, P.F.; Lee, C.-H.; Huang, P.-N.; Tseng, S.-N.; Shih, S.-R.; Huang, H.-Y.; Leu, S.-J.; Huang, Y.-J.; Chiang, L.-C.; Mao, Y.-C.; Wang, W.-C.; Yang, Y.-Y. In Vitro Characterization of Neutralizing Hen Antibodies to Coxsackievirus A16. Int. J. Mol. Sci. 2021, 22, 4146. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084146

AMA Style

Mwale PF, Lee C-H, Huang P-N, Tseng S-N, Shih S-R, Huang H-Y, Leu S-J, Huang Y-J, Chiang L-C, Mao Y-C, Wang W-C, Yang Y-Y. In Vitro Characterization of Neutralizing Hen Antibodies to Coxsackievirus A16. International Journal of Molecular Sciences. 2021; 22(8):4146. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084146

Chicago/Turabian Style

Mwale, Pharaoh F., 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, and Yi-Yuan Yang. 2021. "In Vitro Characterization of Neutralizing Hen Antibodies to Coxsackievirus A16" International Journal of Molecular Sciences 22, no. 8: 4146. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms22084146

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