Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo.
Venezuelan equine encephalitis virus (VEEV) belongs to the Alphavirus genus and several species of this family are pathogenic to humans. The viruses are classified as potential agents of biological warfare and terrorism and sensitive detection as well as effective prophylaxis and antiviral therapies...
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0037242&type=printable |
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| author | Torsten Rülker Luzie Voß Philippe Thullier Lyn M O' Brien Thibaut Pelat Stuart D Perkins Claudia Langermann Thomas Schirrmann Stefan Dübel Hans-Jürgen Marschall Michael Hust Birgit Hülseweh |
| author_facet | Torsten Rülker Luzie Voß Philippe Thullier Lyn M O' Brien Thibaut Pelat Stuart D Perkins Claudia Langermann Thomas Schirrmann Stefan Dübel Hans-Jürgen Marschall Michael Hust Birgit Hülseweh |
| author_sort | Torsten Rülker |
| collection | DOAJ |
| description | Venezuelan equine encephalitis virus (VEEV) belongs to the Alphavirus genus and several species of this family are pathogenic to humans. The viruses are classified as potential agents of biological warfare and terrorism and sensitive detection as well as effective prophylaxis and antiviral therapies are required.In this work, we describe the isolation of the anti-VEEV single chain Fragment variable (scFv), ToR67-3B4, from a non-human primate (NHP) antibody gene library. We report its recloning into the bivalent scFv-Fc format and further immunological and biochemical characterisation.The scFv-Fc ToR67-3B4 recognised viable as well as formalin and ß-propionolactone (ß-Pl) inactivated virus particles and could be applied for immunoblot analysis of VEEV proteins and immuno-histochemistry of VEEV infected cells. It detected specifically the viral E1 envelope protein of VEEV but did not react with reduced viral glycoprotein preparations suggesting that recognition depends upon conformational epitopes. The recombinant antibody was able to detect multiple VEEV subtypes and displayed only marginal cross-reactivity to other Alphavirus species except for EEEV. In addition, the scFv-Fc fusion described here might be of therapeutic use since it successfully inactivated VEEV in a murine disease model. When the recombinant antibody was administered 6 hours post challenge, 80% to 100% of mice survived lethal VEEV IA/B or IE infection. Forty to sixty percent of mice survived when scFv-Fc ToR67-3B4 was applied 6 hours post challenge with VEEV subtypes II and former IIIA. In combination with E2-neutralising antibodies the NHP antibody isolated here could significantly improve passive protection as well as generic therapy of VEE. |
| format | Article |
| id | doaj-art-4d832b7f19f94e06946b856248f9d5d8 |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-4d832b7f19f94e06946b856248f9d5d82025-08-20T03:46:19ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0175e3724210.1371/journal.pone.0037242Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo.Torsten RülkerLuzie VoßPhilippe ThullierLyn M O' BrienThibaut PelatStuart D PerkinsClaudia LangermannThomas SchirrmannStefan DübelHans-Jürgen MarschallMichael HustBirgit HülsewehVenezuelan equine encephalitis virus (VEEV) belongs to the Alphavirus genus and several species of this family are pathogenic to humans. The viruses are classified as potential agents of biological warfare and terrorism and sensitive detection as well as effective prophylaxis and antiviral therapies are required.In this work, we describe the isolation of the anti-VEEV single chain Fragment variable (scFv), ToR67-3B4, from a non-human primate (NHP) antibody gene library. We report its recloning into the bivalent scFv-Fc format and further immunological and biochemical characterisation.The scFv-Fc ToR67-3B4 recognised viable as well as formalin and ß-propionolactone (ß-Pl) inactivated virus particles and could be applied for immunoblot analysis of VEEV proteins and immuno-histochemistry of VEEV infected cells. It detected specifically the viral E1 envelope protein of VEEV but did not react with reduced viral glycoprotein preparations suggesting that recognition depends upon conformational epitopes. The recombinant antibody was able to detect multiple VEEV subtypes and displayed only marginal cross-reactivity to other Alphavirus species except for EEEV. In addition, the scFv-Fc fusion described here might be of therapeutic use since it successfully inactivated VEEV in a murine disease model. When the recombinant antibody was administered 6 hours post challenge, 80% to 100% of mice survived lethal VEEV IA/B or IE infection. Forty to sixty percent of mice survived when scFv-Fc ToR67-3B4 was applied 6 hours post challenge with VEEV subtypes II and former IIIA. In combination with E2-neutralising antibodies the NHP antibody isolated here could significantly improve passive protection as well as generic therapy of VEE.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0037242&type=printable |
| spellingShingle | Torsten Rülker Luzie Voß Philippe Thullier Lyn M O' Brien Thibaut Pelat Stuart D Perkins Claudia Langermann Thomas Schirrmann Stefan Dübel Hans-Jürgen Marschall Michael Hust Birgit Hülseweh Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. PLoS ONE |
| title | Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. |
| title_full | Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. |
| title_fullStr | Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. |
| title_full_unstemmed | Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. |
| title_short | Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. |
| title_sort | isolation and characterisation of a human like antibody fragment scfv that inactivates veev in vitro and in vivo |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0037242&type=printable |
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