Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications
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 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 equivalent to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of live virus replication 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, high-throughput, 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.
| Medienart: |
E-Artikel |
|---|
| Erscheinungsjahr: |
2020 |
|---|---|
| Erschienen: |
2020 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:14 |
|---|---|
| Enthalten in: |
PLoS neglected tropical diseases - 14(2020), 11 vom: 18. Nov., Seite e0008730 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Whitbeck, J Charles [VerfasserIn] |
|---|
| Links: |
|---|
| Themen: |
Antibodies, Monoclonal |
|---|
| Anmerkungen: |
Date Completed 25.01.2021 Date Revised 25.01.2021 published: Electronic-eCollection Citation Status MEDLINE |
|---|
| doi: |
10.1371/journal.pntd.0008730 |
|---|
| funding: |
|
|---|---|
| Förderinstitution / Projekttitel: |
|
| PPN (Katalog-ID): |
NLM317738364 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM317738364 | ||
| 003 | DE-627 | ||
| 005 | 20231225163910.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 231225s2020 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.1371/journal.pntd.0008730 |2 doi | |
| 028 | 5 | 2 | |a pubmed24n1059.xml |
| 035 | |a (DE-627)NLM317738364 | ||
| 035 | |a (NLM)33206639 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Whitbeck, J Charles |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Antigenicity, stability, and reproducibility of Zika reporter virus particles for long-term applications |
| 264 | 1 | |c 2020 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ƒaComputermedien |b c |2 rdamedia | ||
| 338 | |a ƒa Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Date Completed 25.01.2021 | ||
| 500 | |a Date Revised 25.01.2021 | ||
| 500 | |a published: Electronic-eCollection | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 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 equivalent to live virus based on binding ELISA and neutralization results and are nonreplicating based on the results of live virus replication 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, high-throughput, 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 | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Validation Study | |
| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a Antibodies, Neutralizing |2 NLM | |
| 650 | 7 | |a Antibodies, Viral |2 NLM | |
| 650 | 7 | |a Viral Vaccines |2 NLM | |
| 700 | 1 | |a Thomas, Anu |e verfasserin |4 aut | |
| 700 | 1 | |a Kadash-Edmondson, Kathryn |e verfasserin |4 aut | |
| 700 | 1 | |a Grinyo-Escuer, Ariadna |e verfasserin |4 aut | |
| 700 | 1 | |a Stafford, Lewis J |e verfasserin |4 aut | |
| 700 | 1 | |a Cheng, Celine |e verfasserin |4 aut | |
| 700 | 1 | |a Liao, Grant C |e verfasserin |4 aut | |
| 700 | 1 | |a Holtsberg, Frederick W |e verfasserin |4 aut | |
| 700 | 1 | |a Aman, M Javad |e verfasserin |4 aut | |
| 700 | 1 | |a Simmons, Graham |e verfasserin |4 aut | |
| 700 | 1 | |a Davidson, Edgar |e verfasserin |4 aut | |
| 700 | 1 | |a Doranz, Benjamin J |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t PLoS neglected tropical diseases |d 2007 |g 14(2020), 11 vom: 18. Nov., Seite e0008730 |w (DE-627)NLM17489287X |x 1935-2735 |7 nnns |
| 773 | 1 | 8 | |g volume:14 |g year:2020 |g number:11 |g day:18 |g month:11 |g pages:e0008730 |
| 856 | 4 | 0 | |u http://dx.doi.org/10.1371/journal.pntd.0008730 |3 Volltext |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 14 |j 2020 |e 11 |b 18 |c 11 |h e0008730 | ||