A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes

Autores
Depetris, M.; Casalis, P.; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Oggero Eberhardt, Marcos Rafael
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Despite their significant role in maintaining the normal physiology, cytokines may cause pathological conditions when they are overproduced. In this way, the increased expression of human interferon alpha (hIFN-α) is associated with acute viral infections, inflammatory disorders and several autoimmune illnesses, where the cytokine may be a factor in either initiating or maintaining the disease. Currently, there are several mAbs marketed for a variety of indications and many more in clinical trials, in which IFN-α represents a potential target for antibody-based therapy. A panel of 11 murine mAbs was prepared using recombinant hIFN-α2b as immunogen, all of which bound to the native form of the cytokine with affinity constants ranging from 1.7 × 107 M- 1 to 1.4 × 1010 M- 1. An epitope mapping protocol demonstrated four spatially distinct areas of the protein recognized by the mAbs. Taking into account the characterization of the antibodies and their ability to inhibit the IFN-α biological activity, four mAbs were selected to produce scFv fragments. One of these fragments (CA5E6) was able to neutralize a wide spectrum of subtypes of the IFN-α family, including the recombinant cytokines hIFN-α2a and hIFN-α2b and a heterogeneous collection of IFN-α produced by activated leukocytes and Namalwa cells. With the aim of improving the affinity of the selected fragment, a standard error-prone PCR method was carried out. By using this strategy, it was possible to generate a new fragment (EP18) with increased affinity and ability to neutralize a broad diversity of IFN-α subtypes. Consequently, the scFv EP18 represents a potential therapeutic agent for those immune and inflammatory diseases which are associated with an increased IFN-α expression.
Fil: Depetris, M.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Fil: Casalis, P.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Fil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Oggero Eberhardt, Marcos Rafael. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Materia
AUTOIMMUNE DISEASES
INTERFERON ALPHA-2B
NEUTRALIZING ANTIBODY
SCFV
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/114121
Acceder
id CONICETDig_eee3435ddcc62f2ade2b59c686fcc302
oai_identifier_str oai:ri.conicet.gov.ar:11336/114121
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypesDepetris, M.Casalis, P.Kratje, Ricardo BertoldoEtcheverrigaray, MarinaOggero Eberhardt, Marcos RafaelAUTOIMMUNE DISEASESINTERFERON ALPHA-2BNEUTRALIZING ANTIBODYSCFVhttps://purl.org/becyt/ford/3.4https://purl.org/becyt/ford/3Despite their significant role in maintaining the normal physiology, cytokines may cause pathological conditions when they are overproduced. In this way, the increased expression of human interferon alpha (hIFN-α) is associated with acute viral infections, inflammatory disorders and several autoimmune illnesses, where the cytokine may be a factor in either initiating or maintaining the disease. Currently, there are several mAbs marketed for a variety of indications and many more in clinical trials, in which IFN-α represents a potential target for antibody-based therapy. A panel of 11 murine mAbs was prepared using recombinant hIFN-α2b as immunogen, all of which bound to the native form of the cytokine with affinity constants ranging from 1.7 × 107 M- 1 to 1.4 × 1010 M- 1. An epitope mapping protocol demonstrated four spatially distinct areas of the protein recognized by the mAbs. Taking into account the characterization of the antibodies and their ability to inhibit the IFN-α biological activity, four mAbs were selected to produce scFv fragments. One of these fragments (CA5E6) was able to neutralize a wide spectrum of subtypes of the IFN-α family, including the recombinant cytokines hIFN-α2a and hIFN-α2b and a heterogeneous collection of IFN-α produced by activated leukocytes and Namalwa cells. With the aim of improving the affinity of the selected fragment, a standard error-prone PCR method was carried out. By using this strategy, it was possible to generate a new fragment (EP18) with increased affinity and ability to neutralize a broad diversity of IFN-α subtypes. Consequently, the scFv EP18 represents a potential therapeutic agent for those immune and inflammatory diseases which are associated with an increased IFN-α expression.Fil: Depetris, M.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Casalis, P.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaFil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Oggero Eberhardt, Marcos Rafael. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; ArgentinaElsevier Science2008-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/114121Depetris, M.; Casalis, P.; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Oggero Eberhardt, Marcos Rafael; A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes; Elsevier Science; Journal Of Immunological Methods; 334; 1-2; 5-2008; 104-1130022-1759CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jim.2008.02.003info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:47:18Zoai:ri.conicet.gov.ar:11336/114121instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:47:18.612CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
title A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
spellingShingle A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
Depetris, M.
AUTOIMMUNE DISEASES
INTERFERON ALPHA-2B
NEUTRALIZING ANTIBODY
SCFV
title_short A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
title_full A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
title_fullStr A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
title_full_unstemmed A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
title_sort A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes
dc.creator.none.fl_str_mv Depetris, M.
Casalis, P.
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
Oggero Eberhardt, Marcos Rafael
author Depetris, M.
author_facet Depetris, M.
Casalis, P.
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
Oggero Eberhardt, Marcos Rafael
author_role author
author2 Casalis, P.
Kratje, Ricardo Bertoldo
Etcheverrigaray, Marina
Oggero Eberhardt, Marcos Rafael
author2_role author
author
author
author
dc.subject.none.fl_str_mv AUTOIMMUNE DISEASES
INTERFERON ALPHA-2B
NEUTRALIZING ANTIBODY
SCFV
topic AUTOIMMUNE DISEASES
INTERFERON ALPHA-2B
NEUTRALIZING ANTIBODY
SCFV
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.4
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Despite their significant role in maintaining the normal physiology, cytokines may cause pathological conditions when they are overproduced. In this way, the increased expression of human interferon alpha (hIFN-α) is associated with acute viral infections, inflammatory disorders and several autoimmune illnesses, where the cytokine may be a factor in either initiating or maintaining the disease. Currently, there are several mAbs marketed for a variety of indications and many more in clinical trials, in which IFN-α represents a potential target for antibody-based therapy. A panel of 11 murine mAbs was prepared using recombinant hIFN-α2b as immunogen, all of which bound to the native form of the cytokine with affinity constants ranging from 1.7 × 107 M- 1 to 1.4 × 1010 M- 1. An epitope mapping protocol demonstrated four spatially distinct areas of the protein recognized by the mAbs. Taking into account the characterization of the antibodies and their ability to inhibit the IFN-α biological activity, four mAbs were selected to produce scFv fragments. One of these fragments (CA5E6) was able to neutralize a wide spectrum of subtypes of the IFN-α family, including the recombinant cytokines hIFN-α2a and hIFN-α2b and a heterogeneous collection of IFN-α produced by activated leukocytes and Namalwa cells. With the aim of improving the affinity of the selected fragment, a standard error-prone PCR method was carried out. By using this strategy, it was possible to generate a new fragment (EP18) with increased affinity and ability to neutralize a broad diversity of IFN-α subtypes. Consequently, the scFv EP18 represents a potential therapeutic agent for those immune and inflammatory diseases which are associated with an increased IFN-α expression.
Fil: Depetris, M.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Fil: Casalis, P.. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
Fil: Kratje, Ricardo Bertoldo. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Etcheverrigaray, Marina. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Oggero Eberhardt, Marcos Rafael. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Cultivos Celulares; Argentina
description Despite their significant role in maintaining the normal physiology, cytokines may cause pathological conditions when they are overproduced. In this way, the increased expression of human interferon alpha (hIFN-α) is associated with acute viral infections, inflammatory disorders and several autoimmune illnesses, where the cytokine may be a factor in either initiating or maintaining the disease. Currently, there are several mAbs marketed for a variety of indications and many more in clinical trials, in which IFN-α represents a potential target for antibody-based therapy. A panel of 11 murine mAbs was prepared using recombinant hIFN-α2b as immunogen, all of which bound to the native form of the cytokine with affinity constants ranging from 1.7 × 107 M- 1 to 1.4 × 1010 M- 1. An epitope mapping protocol demonstrated four spatially distinct areas of the protein recognized by the mAbs. Taking into account the characterization of the antibodies and their ability to inhibit the IFN-α biological activity, four mAbs were selected to produce scFv fragments. One of these fragments (CA5E6) was able to neutralize a wide spectrum of subtypes of the IFN-α family, including the recombinant cytokines hIFN-α2a and hIFN-α2b and a heterogeneous collection of IFN-α produced by activated leukocytes and Namalwa cells. With the aim of improving the affinity of the selected fragment, a standard error-prone PCR method was carried out. By using this strategy, it was possible to generate a new fragment (EP18) with increased affinity and ability to neutralize a broad diversity of IFN-α subtypes. Consequently, the scFv EP18 represents a potential therapeutic agent for those immune and inflammatory diseases which are associated with an increased IFN-α expression.
publishDate 2008
dc.date.none.fl_str_mv 2008-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/114121
Depetris, M.; Casalis, P.; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Oggero Eberhardt, Marcos Rafael; A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes; Elsevier Science; Journal Of Immunological Methods; 334; 1-2; 5-2008; 104-113
0022-1759
CONICET Digital
CONICET
url http://hdl.handle.net/11336/114121
identifier_str_mv Depetris, M.; Casalis, P.; Kratje, Ricardo Bertoldo; Etcheverrigaray, Marina; Oggero Eberhardt, Marcos Rafael; A scFv antibody fragment as a therapeutic candidate to neutralize a broad diversity of human IFN-alpha subtypes; Elsevier Science; Journal Of Immunological Methods; 334; 1-2; 5-2008; 104-113
0022-1759
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jim.2008.02.003
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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