In Vitro Assembly and Stabilization of Dengue and Zika Virus Envelope Protein Homo-Dimers

Metz, Stefan; Gallichotte, Emily N.; Brackbill, Alex; Premkumar, Lakshmanane; Miley, Michael J.; Baric, Ralph S.; Silva, Aravinda M. de

doi:10.1038/s41598-017-04767-6

Cited by 41 publications

(48 citation statements)

References 41 publications

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“…ZIKV recombinant E protein was purified as described previously (82) and conjugated to HisPur Ni-NTA magnetic beads (Thermo Fisher Scientific) per manufacturer's instructions. Control beads were incubated with an equal amount His-tagged human MBP (His-MBP).…”

Section: Ab Depletionsmentioning

confidence: 99%

Human antibody response to Zika targets type-specific quaternary structure epitopes

Collins¹,

Tu²,

Gimblet-Ochieng³

et al. 2019

JCI Insight

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Section: Ab Depletionsmentioning

confidence: 99%

Human antibody response to Zika targets type-specific quaternary structure epitopes

Collins¹,

Tu²,

Gimblet-Ochieng³

et al. 2019

JCI Insight

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“…Maltose binding protein (MBP) fused to Z-EDI or Z-EDIII was expressed in Escherichia coli BL21(DE3)pLysS and purified using amylose affinity resin. The ZIKV E80 antigen (aa 1 to 404) was expressed in the Expi293 transient expression system and purified by use of Ni-nitrilotriacetic acid (Ni-NTA) affinity resin as previously described (30,31).…”

mentioning

confidence: 99%

Development of Envelope Protein Antigens To Serologically Differentiate Zika Virus Infection from Dengue Virus Infection

et al. 2018

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Zika virus (ZIKV) is an emerging flavivirus that can cause birth defects and neurologic complications. Molecular tests are effective for diagnosing acute ZIKV infection, although the majority of infections produce no symptoms at all or present after the narrow window in which molecular diagnostics are dependable. Serology is a reliable method for detecting infections after the viremic period; however, most serological assays have limited specificity due to cross-reactive antibodies elicited by flavivirus infections. Since ZIKV and dengue virus (DENV) widely cocirculate, distinguishing ZIKV infection from DENV infection is particularly important for diagnosing individual cases or for surveillance to coordinate public health responses. Flaviviruses also elicit type-specific antibodies directed to non-cross-reactive epitopes of the infecting virus; such epitopes are attractive targets for the design of antigens for development of serological tests with greater specificity. Guided by comparative epitope modeling of the ZIKV envelope protein, we designed two recombinant antigens displaying unique antigenic regions on domain I (Z-EDI) and domain III (Z-EDIII) of the ZIKV envelope protein. Both the Z-EDI and Z-EDIII antigens consistently detected ZIKV-specific IgG in ZIKV-immune sera but not cross-reactive IgG in DENV-immune sera in late convalescence (>12 weeks postinfection). In contrast, during early convalescence (2 to 12 weeks postinfection), secondary DENV-immune sera and some primary DENV-immune sera cross-reacted with the Z-EDI and Z-EDIII antigens. Analysis of sequential samples from DENV-immune individuals demonstrated that Z-EDIII cross-reactivity peaked in early convalescence and declined steeply over time. The Z-EDIII antigen has much potential as a diagnostic antigen for population-level surveillance and for detecting past infections in patients.

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“…dengue virus E, etc. ), whose glycoproteins are frequently difficult to produce in recombinant form (35). Incorporation of sustainable and sensitive immunoassay reagents such as VCMs into field-forward technologies will become increasingly important in global surveillance and diagnostic efforts to limit the spread of infectious viral diseases.…”

Section: Discussionmentioning

confidence: 99%

Virus-Like Particles and Magnetic Microspheres Provide a Flexible and Sustainable Multiplexed Alphavirus Immunodiagnostic Platform

Ricks¹,

Shoemaker

Dupuy³

et al. 2018

Preprint

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14There is a pressing need for sustainable and sensitive immunodiagnostics for use in public health 15 efforts to understand and combat the threat of endemic and emerging infectious diseases. We 16 describe a novel approach to immunodiagnostics based on virus-like particles (VLPs) attached to 17 magnetic beads. This flexible, innovative immunoassay system, based on the MAGPIX® 18 platform, improves sensitivity by up to 2-logs and has faster sample-to-answer time over 19 traditional methods. As a proof of concept, a retroviral-based VLP, that presents the Venezuelan 20 equine encephalitis virus E1/E2 glycoprotein antigen on its surface, was generated and coupled 21 to magnetic beads to create VLP-conjugated microspheres (VCMs). Using these VCMs, IgG and 22IgM antibodies were detectable in nonhuman primate (NHP) and human clinical serum samples 23 at dilutions of 1 × 10 4 and greater. We extended the VCM methodology to two other New-World 24 alphaviruses, eastern and western equine encephalitis viruses, as well as an Old-World 25 alphavirus, Chikungunya virus, demonstrating the flexibility of this approach toward different 26 VLP architectures. When multiplexed on the MAGPIX platform, the VCMs provided 27 differential diagnosis between Old-World and New-World alphaviruses and well as a route 28 toward assessing the humoral response to both natural infection and vaccination. This VCM 29 system will allow more rapid and efficient detection of endemic and emerging viral pathogens in 30 human populations. 31

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In Vitro Assembly and Stabilization of Dengue and Zika Virus Envelope Protein Homo-Dimers

Cited by 41 publications

References 41 publications

Human antibody response to Zika targets type-specific quaternary structure epitopes

Human antibody response to Zika targets type-specific quaternary structure epitopes

Development of Envelope Protein Antigens To Serologically Differentiate Zika Virus Infection from Dengue Virus Infection

Virus-Like Particles and Magnetic Microspheres Provide a Flexible and Sustainable Multiplexed Alphavirus Immunodiagnostic Platform

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