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Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications

J. Charles Whitbeck, Anu Thomas, Kathryn Kadash-Edmondson, Ariadna Grinyo i Escuer, Celine Cheng, Grant C. Liao, Frederick W. Holtsberg, M. Javad Aman, Graham Simmons, Edgar Davidson, Benjamin J. Doranz
doi: https://doi.org/10.1101/2020.04.21.047241
J. Charles Whitbeck
1Integral Molecular, Inc., 3711 Market St, Philadelphia, PA 19104
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Anu Thomas
1Integral Molecular, Inc., 3711 Market St, Philadelphia, PA 19104
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Kathryn Kadash-Edmondson
1Integral Molecular, Inc., 3711 Market St, Philadelphia, PA 19104
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Ariadna Grinyo i Escuer
1Integral Molecular, Inc., 3711 Market St, Philadelphia, PA 19104
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Celine Cheng
2Vitalant Research Institute, 270 Masonic Ave, San Francisco, CA 94118
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Grant C. Liao
3Integrated Biotherapeutics, 4 Research Court, Suite 300 Rockville, MD 20850
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Frederick W. Holtsberg
3Integrated Biotherapeutics, 4 Research Court, Suite 300 Rockville, MD 20850
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M. Javad Aman
3Integrated Biotherapeutics, 4 Research Court, Suite 300 Rockville, MD 20850
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Graham Simmons
2Vitalant Research Institute, 270 Masonic Ave, San Francisco, CA 94118
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Edgar Davidson
1Integral Molecular, Inc., 3711 Market St, Philadelphia, PA 19104
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Benjamin J. Doranz
1Integral Molecular, Inc., 3711 Market St, Philadelphia, PA 19104
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  • For correspondence: bdoranz@integralmolecular.com
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ABSTRACT

The development of vaccines against flaviviruses, including Zika virus (ZIKV) and dengue virus (DENV), continues to be a major challenge, hindered by the lack of efficient and reliable methods for screening neutralizing activity of sera or antibodies. To address this need, we previously developed a plasmid-based, replication-incompetent DENV reporter virus particle (RVP) production system as an efficient and safe alternative to the Plaque Reduction Neutralization Test (PRNT). As part of the response to the 2015-2016 ZIKV outbreak, we recently developed pseudo-infectious ZIKV RVPs by modifying our DENV RVP system. The use of ZIKV RVPs as critical reagents in human clinical trials requires their further validation using stability and reproducibility metrics for large-scale applications. In the current study, we validated ZIKV RVPs using infectivity, neutralization, and enhancement assays with monoclonal antibodies (MAbs) and human ZIKV-positive patient serum. ZIKV RVPs are antigenically identical to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of plaque formation assays. We demonstrate reproducible neutralization titer data (NT50 values) across different RVP production lots, volumes, time frames, and laboratories. We also show RVP stability across experimentally relevant time intervals and temperatures. Our results demonstrate that ZIKV RVPs provide a safe, rapid, and reproducible reagent for large-scale, long-term studies of neutralizing antibodies and sera, which can facilitate large-scale screening and epidemiological studies to help expedite ZIKV vaccine development.

AUTHOR SUMMARY ZIKV is a mosquito-borne virus that can cause severe birth defects and other disorders. Large outbreaks of ZIKV occurred in 2015 and 2016 and there are still no drugs or vaccines available to protect against ZIKV infection. Vaccine development has been hindered by the lack of safe and efficient methods to screen ZIKV neutralizing properties of patient sera or antibodies, especially in the context of large clinical trials. To address this unmet need, we developed and validated the use of ZIKV reporter virus particles (RVPs), a safe, high-throughput and quantitative alternative to using live virus for neutralization studies. We show that ZIKV RVPs are stable, show lot-to-lot consistency and provide reproducible neutralization data that is suitable for large-scale studies are needed for development of a ZIKV vaccine, epidemiologic surveillance, and high-throughput screening.

Competing Interest Statement

Integral Molecular produces and sells ZIKV RVPs as a commercial product. J. Charles Whitbeck, Anu Thomas, Kathryn Kadash-Edmondson, Ariadna Grinyo i Escuer, Edgar Davidson and Benjamin J. Doranz are all current or former employees of Integral Molecular.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted April 22, 2020.
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Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications
J. Charles Whitbeck, Anu Thomas, Kathryn Kadash-Edmondson, Ariadna Grinyo i Escuer, Celine Cheng, Grant C. Liao, Frederick W. Holtsberg, M. Javad Aman, Graham Simmons, Edgar Davidson, Benjamin J. Doranz
bioRxiv 2020.04.21.047241; doi: https://doi.org/10.1101/2020.04.21.047241
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Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications
J. Charles Whitbeck, Anu Thomas, Kathryn Kadash-Edmondson, Ariadna Grinyo i Escuer, Celine Cheng, Grant C. Liao, Frederick W. Holtsberg, M. Javad Aman, Graham Simmons, Edgar Davidson, Benjamin J. Doranz
bioRxiv 2020.04.21.047241; doi: https://doi.org/10.1101/2020.04.21.047241

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