Cargando…
Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3
Foot-and-mouth disease (FMD) affects cloven-hoofed domestic and wildlife animals and an outbreak can cause severe losses in milk production, reduction in meat production and death amongst young animals. Several parts of Asia, most of Africa, and the Middle East remain endemic, thus emphasis on impro...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7432252/ https://www.ncbi.nlm.nih.gov/pubmed/32851044 http://dx.doi.org/10.3389/fvets.2020.00475 |
_version_ | 1783571757162037248 |
---|---|
author | Chitray, Melanie Opperman, Pamela Anne Rotherham, Lia Fehrsen, Jeanni van Wyngaardt, Wouter Frischmuth, Janine Rieder, Elizabeth Maree, Francois Frederick |
author_facet | Chitray, Melanie Opperman, Pamela Anne Rotherham, Lia Fehrsen, Jeanni van Wyngaardt, Wouter Frischmuth, Janine Rieder, Elizabeth Maree, Francois Frederick |
author_sort | Chitray, Melanie |
collection | PubMed |
description | Foot-and-mouth disease (FMD) affects cloven-hoofed domestic and wildlife animals and an outbreak can cause severe losses in milk production, reduction in meat production and death amongst young animals. Several parts of Asia, most of Africa, and the Middle East remain endemic, thus emphasis on improved FMD vaccines, diagnostic assays, and control measures are key research areas. FMD virus (FMDV) populations are quasispecies, which pose serious implications in vaccine design and efficacy where an effective vaccine should include multiple independent neutralizing epitopes to elicit an adequate immune response. Further investigation of the residues that comprise the antigenic determinants of the virus will allow the identification of mutations in outbreak strains that potentially lessen the efficacy of a vaccine. Additionally, of utmost importance in endemic regions, is the accurate diagnosis of FMDV infection for the control and eradication of the disease. To this end, a phage display library was explored to identify FMDV epitopes for recombinant vaccines and for the generation of reagents for improved diagnostic FMD enzyme-linked immunosorbent assays (ELISAs). A naïve semi-synthetic chicken single chain variable fragment (scFv) phage display library i.e., the Nkuku(®) library was used for bio-panning against FMD Southern-African Territories (SAT) 1, SAT3, and serotype A viruses. Biopanning yielded one unique scFv against SAT1, two for SAT3, and nine for A22. SAT1 and SAT3 specific scFvs were exploited as capturing and detecting reagents to develop an improved diagnostic ELISA for FMDV. The SAT1 soluble scFv showed potential as a detecting reagent in the liquid phase blocking ELISA (LPBE) as it reacted specifically with a panel of SAT1 viruses, albeit with different ELISA absorbance signals. The SAT1svFv1 had little or no change on its paratope when coated on polystyrene plates whilst the SAT3scFv's paratope may have changed. SAT1 and SAT3 soluble scFvs did not neutralize the SAT1 and SAT3 viruses; however, three of the nine A22 binders i.e., A22scFv1, A22scFv2, and A22scFv8 were able to neutralize A22 virus. Following the generation of virus escape mutants through successive virus passage under scFv pressure, FMDV epitopes were postulated i.e., RGD+3 and +4 positions respectively, proving the epitope mapping potential of scFvs. |
format | Online Article Text |
id | pubmed-7432252 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-74322522020-08-25 Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 Chitray, Melanie Opperman, Pamela Anne Rotherham, Lia Fehrsen, Jeanni van Wyngaardt, Wouter Frischmuth, Janine Rieder, Elizabeth Maree, Francois Frederick Front Vet Sci Veterinary Science Foot-and-mouth disease (FMD) affects cloven-hoofed domestic and wildlife animals and an outbreak can cause severe losses in milk production, reduction in meat production and death amongst young animals. Several parts of Asia, most of Africa, and the Middle East remain endemic, thus emphasis on improved FMD vaccines, diagnostic assays, and control measures are key research areas. FMD virus (FMDV) populations are quasispecies, which pose serious implications in vaccine design and efficacy where an effective vaccine should include multiple independent neutralizing epitopes to elicit an adequate immune response. Further investigation of the residues that comprise the antigenic determinants of the virus will allow the identification of mutations in outbreak strains that potentially lessen the efficacy of a vaccine. Additionally, of utmost importance in endemic regions, is the accurate diagnosis of FMDV infection for the control and eradication of the disease. To this end, a phage display library was explored to identify FMDV epitopes for recombinant vaccines and for the generation of reagents for improved diagnostic FMD enzyme-linked immunosorbent assays (ELISAs). A naïve semi-synthetic chicken single chain variable fragment (scFv) phage display library i.e., the Nkuku(®) library was used for bio-panning against FMD Southern-African Territories (SAT) 1, SAT3, and serotype A viruses. Biopanning yielded one unique scFv against SAT1, two for SAT3, and nine for A22. SAT1 and SAT3 specific scFvs were exploited as capturing and detecting reagents to develop an improved diagnostic ELISA for FMDV. The SAT1 soluble scFv showed potential as a detecting reagent in the liquid phase blocking ELISA (LPBE) as it reacted specifically with a panel of SAT1 viruses, albeit with different ELISA absorbance signals. The SAT1svFv1 had little or no change on its paratope when coated on polystyrene plates whilst the SAT3scFv's paratope may have changed. SAT1 and SAT3 soluble scFvs did not neutralize the SAT1 and SAT3 viruses; however, three of the nine A22 binders i.e., A22scFv1, A22scFv2, and A22scFv8 were able to neutralize A22 virus. Following the generation of virus escape mutants through successive virus passage under scFv pressure, FMDV epitopes were postulated i.e., RGD+3 and +4 positions respectively, proving the epitope mapping potential of scFvs. Frontiers Media S.A. 2020-08-11 /pmc/articles/PMC7432252/ /pubmed/32851044 http://dx.doi.org/10.3389/fvets.2020.00475 Text en Copyright © 2020 Chitray, Opperman, Rotherham, Fehrsen, van Wyngaardt, Frischmuth, Rieder and Maree. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Veterinary Science Chitray, Melanie Opperman, Pamela Anne Rotherham, Lia Fehrsen, Jeanni van Wyngaardt, Wouter Frischmuth, Janine Rieder, Elizabeth Maree, Francois Frederick Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 |
title | Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 |
title_full | Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 |
title_fullStr | Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 |
title_full_unstemmed | Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 |
title_short | Diagnostic and Epitope Mapping Potential of Single-Chain Antibody Fragments Against Foot-and-Mouth Disease Virus Serotypes A, SAT1, and SAT3 |
title_sort | diagnostic and epitope mapping potential of single-chain antibody fragments against foot-and-mouth disease virus serotypes a, sat1, and sat3 |
topic | Veterinary Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7432252/ https://www.ncbi.nlm.nih.gov/pubmed/32851044 http://dx.doi.org/10.3389/fvets.2020.00475 |
work_keys_str_mv | AT chitraymelanie diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT oppermanpamelaanne diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT rotherhamlia diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT fehrsenjeanni diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT vanwyngaardtwouter diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT frischmuthjanine diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT riederelizabeth diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 AT mareefrancoisfrederick diagnosticandepitopemappingpotentialofsinglechainantibodyfragmentsagainstfootandmouthdiseasevirusserotypesasat1andsat3 |