Expression, Purification, and Characterization of Anti-Zika virus Envelope Protein: Polyclonal and Chicken-Derived Single Chain Variable Fragment Antibodies.

Pharaoh Fellow Mwale,Sy-Jye Leu,Liao-Chun Chiang,Yan-Chiao Mao,Chi-Hsin Lee,Yi-Yuan Yang,Liang-Tzung Lin,Yun-Ju Huang

doi:10.3390/ijms21020492

Abstract

Zika virus (ZIKV) is a new and emerging virus that has caused outbreaks worldwide. The virus has been linked to congenital neurological malformations in neonates and Guillain–Barré syndrome in adults. Currently there are no effective vaccines available. As a result, there is a great need for ZIKV treatment. In this study, we developed single chain variable fragment (scFv) antibodies that target the ZIKV envelope protein using phage display technology. We first induced an immune response in white leghorn laying hens against the ZIKV envelope (E) protein. Chickens were immunized and polyclonal immunoglobulin yolk (IgY) antibodies were extracted from egg yolks. A high-level titer of anti-ZIKV_E IgY antibodies was detected using enzyme-linked immunosorbent assay (ELISA) after the third immunization. The titer persisted for at least 9 weeks. We constructed two antibody libraries that contained 5.3 × 106 and 4.5 × 106 transformants. After biopanning, an ELISA phage assay confirmed the enrichment of specific clones. We randomly selected 26 clones that expressed ZIKV scFv antibodies and classified them into two groups, short-linker and long-linker. Of these, four showed specific binding activities toward ZIKV_E proteins. These data suggest that the polyclonal and monoclonal scFv antibodies have the diagnostic or therapeutic potential for ZIKV.

Highlights

Zika virus (ZIKV) is an arthropod-borne virus and a member of the Flavivirus genus belonging to the Flaviviradae family
ZIKV has been associated with several health issues, such as meningoencephalitis and myelitis in adults [6]; autoimmune disorders such as Guillain–Barré syndrome (GBS) in adults [7,8]; and microcephaly, which occurs in fetuses who were born from infected pregnant women [9]
We report the first analysis of the immune response in chicken through determination of the polyclonal immunoglobulin yolk (IgY) against truncated Zika virus envelope (ZIKV_E) protein of the Brazilian lineage ZIKV strain [27]

Summary

Introduction

Zika virus (ZIKV) is an arthropod-borne virus and a member of the Flavivirus genus belonging to the Flaviviradae family. ZIKV has been associated with several health issues, such as meningoencephalitis and myelitis in adults [6]; autoimmune disorders such as Guillain–Barré syndrome (GBS) in adults [7,8]; and microcephaly, which occurs in fetuses who were born from infected pregnant women [9]. Following its outbreak in Brazil, scientists reported an association between ZIKV and microcephaly cases in infants [9]. Research has suggested that individuals with compromised immunity could be more susceptible to ZIKV infection and disease development [11]. Much effort has been made towards preventing and curing ZIKV infections. No vaccines or drugs are currently approved on the market for the disease-causing ZIKV

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Results

Conclusion