An Approach for Zika Virus Inhibition Using Homology Structure of the Envelope Protein

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.

Download Full-text

Dengue virus envelope protein domain III-elicited antibodies mediate cross-protection against Zika virus in a mouse model

Virus Research ◽

10.1016/j.virusres.2020.197882 ◽

2020 ◽

Vol 278 ◽

pp. 197882

Author(s):

Yongchao Zhou ◽

Dong Chen ◽

Lan Yang ◽

Weiwei Zou ◽

Zhiliang Duan ◽

...

Keyword(s):

Mouse Model ◽

Dengue Virus ◽

Zika Virus ◽

Envelope Protein ◽

Cross Protection ◽

Protein Domain ◽

Virus Envelope ◽

Virus Envelope Protein ◽

Domain Iii

Download Full-text

Structures of the Zika Virus Envelope Protein and Its Complex with a Flavivirus Broadly Protective Antibody

Cell Host & Microbe ◽

10.1016/j.chom.2016.04.013 ◽

2016 ◽

Vol 19 (5) ◽

pp. 696-704 ◽

Cited By ~ 277

Author(s):

Lianpan Dai ◽

Jian Song ◽

Xishan Lu ◽

Yong-Qiang Deng ◽

Abednego Moki Musyoki ◽

...

Keyword(s):

Zika Virus ◽

Envelope Protein ◽

Virus Envelope ◽

Protective Antibody ◽

Virus Envelope Protein

Download Full-text

Antivirals for allosteric inhibition of Zika virus using a homology model and experimentally determined structure of envelope protein

BMC Research Notes ◽

10.1186/s13104-017-2685-7 ◽

2017 ◽

Vol 10 (1) ◽

Cited By ~ 3

Author(s):

Sandun Fernando ◽

Teshan Fernando

Keyword(s):

Zika Virus ◽

Envelope Protein ◽

Homology Model ◽

Allosteric Inhibition

Download Full-text

Functional activity of antisera against recombinant Zika virus envelope protein subunits expressed in Escherichia coli

10.1101/698266 ◽

2019 ◽

Author(s):

Hong-Yun Tham ◽

Man Kwan Ooi ◽

Vinod RMT Balasubramaniam ◽

Sharifah Syed Hassan ◽

Hong-Wai Tham

Keyword(s):

Zika Virus ◽

Envelope Protein ◽

Mammalian Cells ◽

Vaccine Development ◽

Vaccine Candidate ◽

Cross Reactivity ◽

Virus Envelope ◽

Humoral And Cellular Immunity ◽

Progressive Development

AbstractThe global Zika virus (ZIKV) outbreak across continents has been drawing research attentions to researchers and healthcare professionals. It highlights the urgent development of ZIKV vaccines that offer rapid, precise and specific protection to those living in the high-risk regions - the tropical and subtropical regions. As a public health priority, there is a progressive development in the discovery of vaccine candidates and design in recent years. Many efforts have been placed in the in vitro development of ZIKV subunits as the vaccine candidate in various protein expression systems, including bacteria, yeast, plant cells, insect cells and mammalian cells. However, due to the lack of knowledge on humoral and cellular immune responses against virus vaccines, a commercialised vaccine against Dengue virus (DENV) has been suspended due to a health scare in Philippines. Moreover, the closely-related DENV and ZIKV has indicated serological cross-reactivity between both viruses. This has led to greater attentions to precautions needed during the design of ZIKV and DENV vaccines. In this study, we pre-selected, synthesised and expressed the domain III of ZIKV envelope protein (namely rEDIII) based on a previously-established report (GenBank: AMC13911.1). The characteristics of purified ZIKV rEDIII was tested using SDS-PAGE, Western blotting and LC-MS/MS. Since the ZIKV rEDIII has been well reported as a potential protein candidate in ZIKV vaccine development, we assessed the possible outcome of preexisting immunity against the rEDIII proteins by conducting dot-blotting assays using mice antisera pre-immunised with ZIKV particles (ZIKV strain: MRS_OPY_Martinique_PaRi_2015, GenBank: KU647676) . Surprisingly, the antisera was able to recognise the rEDIII of a different ZIKV strain (GenBank: AMC13911.1). Despite its great antigenicity in eliciting humoral and cellular immunity against ZIKV infection, our finding calls for greater attention to evaluate the details of ZIKV rEDIII as a stand-alone vaccine candidate.

Download Full-text

Zika virus baculovirus-expressed envelope protein elicited humoral and cellular immunity in immunocompetent mice

Scientific Reports ◽

10.1038/s41598-021-04713-7 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Minna Shin ◽

Kiju Kim ◽

Hyo-Ji Lee ◽

Rangyeon Lee ◽

Yu-Jin Jung ◽

...

Keyword(s):

Zika Virus ◽

Envelope Protein ◽

Expression System ◽

Baculovirus Expression System ◽

Interferon Γ ◽

Interleukin 12 ◽

Effective Vaccine ◽

Vaccine Strategy ◽

Humoral And Cellular Immunity ◽

Baculovirus Expression

AbstractZika virus (ZIKV) is a mosquito-borne virus that has a high risk of inducing Guillain–Barré syndrome and microcephaly in newborns. Because vaccination is considered the most effective strategy against ZIKV infection, we designed a recombinant vaccine utilizing the baculovirus expression system with two strains of ZIKV envelope protein (MR766, Env_M; ZBRX6, Env_Z). Animals inoculated with Env_M and Env_Z produced ZIKV-specific antibodies and secreted effector cytokines such as interferon-γ, tumor necrosis factor-α, and interleukin-12. Moreover, the progeny of immunized females had detectable maternal antibodies that protected them against two ZIKV strains (MR766 and PRVABC59) and a Dengue virus strain. We propose that the baculovirus expression system ZIKV envelope protein recombinant provides a safe and effective vaccine strategy.

Download Full-text

A global map of the Zika virus phosphoproteome reveals host-driven regulation of viral budding

10.1101/2021.11.24.469793 ◽

2021 ◽

Author(s):

Inessa Manuelyan ◽

Anna M. Schmoker ◽

Boyd L. Yount ◽

Philip Eisenhauer ◽

Judith I. Keller ◽

...

Keyword(s):

Zika Virus ◽

Envelope Protein ◽

Human Population ◽

Kinase Inhibitor ◽

Health Crisis ◽

Post Translational Modifications ◽

Viral Propagation ◽

Viral Budding ◽

High Resolution Map ◽

Positive Strand Rna

Flaviviruses are enveloped, positive-strand RNA viruses that cause millions of infections in the human population annually. Although Zika virus (ZIKV) had been detected in humans as early as the 1950s, its reemergence in South America in 2015 resulted in a global health crisis. While flaviviruses encode 10 proteins that can be post-translationally modified by host enzymes, little is known regarding post-translational modifications (PTMs) of the flavivirus proteome. We used mass spectrometry to comprehensively identify host-driven PTMs on the ZIKV proteome. This approach allowed us to identify 43 PTMs across 8 ZIKV proteins, including several that are highly conserved within the Flavivirus genus. Notably, we found two phosphosites on the ZIKV envelope protein that are functionally important for viral propagation and appear to regulate viral budding. Additionally, we discovered 115 host kinases that interacted with ZIKV proteins and determined that Bosutinib, an FDA-approved tyrosine kinase inhibitor that targets ZIKV interacting host kinases, impairs ZIKV growth. Thus, we have defined a high-resolution map of host-driven PTMs on ZIKV proteins as well as cellular interacting kinases, uncovered a novel mechanism of host driven-regulation of ZIKV budding, and identified an FDA-approved inhibitor of ZIKV growth.

Download Full-text