North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens

We report the cloning of sphingomyelinase D (SMD) cDNA from Loxosceles reclusa, Loxosceles boneti and Loxosceles laeta into bacterial expression systems, as well as optimization of expression conditions so as to obtain soluble and active recombinant enzymes. The recombinant mature SMDs, tagged with...

Descripción completa

Detalles Bibliográficos
Autores: Olvera, A, Ramos-Cerrillo, B, Estevez, J, Clement, H, de Roodt, A, Paniagua-Solis, J, Vazquez, H, Zavaleta, A, Arruz, MS, Stock, RP, Alagon, A
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2006
País:México
Institución:Universidad Nacional Autónoma de México
Repositorio:Sistema de Información de la Facultad de Ciencias, UNAM
OAI Identifier:oai:repositorio.fciencias.unam.mx:11154/3190
Acceso en línea:http://hdl.handle.net/11154/3190
Access Level:acceso abierto
Palabra clave:Pharmacology & Pharmacy
Toxicology
Loxosceles
recombinant sphingomyelinase D
enzymatic activity
antivenom
id MX_5cc335aab7eebd50d89e94fc5cb2f2a9
oai_identifier_str oai:repositorio.fciencias.unam.mx:11154/3190
network_acronym_str MX
network_name_str México
repository_id_str
spelling North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigensOlvera, ARamos-Cerrillo, BEstevez, JClement, Hde Roodt, APaniagua-Solis, JVazquez, HZavaleta, AArruz, MSStock, RPAlagon, APharmacology & PharmacyToxicologyLoxoscelesrecombinant sphingomyelinase Denzymatic activityantivenomWe report the cloning of sphingomyelinase D (SMD) cDNA from Loxosceles reclusa, Loxosceles boneti and Loxosceles laeta into bacterial expression systems, as well as optimization of expression conditions so as to obtain soluble and active recombinant enzymes. The recombinant mature SMDs, tagged with a histidine tail at the N- or C-termini, were compared in terms of toxicity and enzymatic activity, and were used as immunogens for the production of monovalent antiscra in rabbits and F(ab')(2) preparations in animals used for commercial antivenom production (horses). We performed studies on in vitro inhibition of enzymatic activity of natural venom preparations by antibodies generated against the tagged proteins. We also present and discuss the results of studies on the specific and para-specific in vivo protective potential of the rabbit and equine antibody preparations against the recombinant proteins themselves and natural venom preparations. Our conclusions support the feasibility of using recombinant SMDs for production and evaluation of polyvalent anti-Loxosceles antivenoms, and we offer data on the potential of paraspecific neutralization in the context of the antigenic groupings and the molecular phylogeny of those active SMDs for which amino acid sequence information is available. (c) 2006 Elsevier Ltd. All rights reserved.2011-01-22T10:26:24Z2011-01-22T10:26:24Z2006info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article0041-0101http://hdl.handle.net/11154/3190134810.1016/j.toxicon.2006.04.01048(1):64-74reponame:Sistema de Información de la Facultad de Ciencias, UNAMinstname:Universidad Nacional Autónoma de Méxicoinstacron:UNAMenToxiconinfo:eu-repo/semantics/openAccessoai:repositorio.fciencias.unam.mx:11154/31902025-09-17T19:21:34Z
dc.title.none.fl_str_mv North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
title North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
spellingShingle North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
Olvera, A
Pharmacology & Pharmacy
Toxicology
Loxosceles
recombinant sphingomyelinase D
enzymatic activity
antivenom
title_short North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
title_full North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
title_fullStr North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
title_full_unstemmed North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
title_sort North and South American Loxosceles spiders: Development of a polyvalent antivenom with recombinant sphingomyelinases D as antigens
dc.creator.none.fl_str_mv Olvera, A
Ramos-Cerrillo, B
Estevez, J
Clement, H
de Roodt, A
Paniagua-Solis, J
Vazquez, H
Zavaleta, A
Arruz, MS
Stock, RP
Alagon, A
author Olvera, A
author_facet Olvera, A
Ramos-Cerrillo, B
Estevez, J
Clement, H
de Roodt, A
Paniagua-Solis, J
Vazquez, H
Zavaleta, A
Arruz, MS
Stock, RP
Alagon, A
author_role author
author2 Ramos-Cerrillo, B
Estevez, J
Clement, H
de Roodt, A
Paniagua-Solis, J
Vazquez, H
Zavaleta, A
Arruz, MS
Stock, RP
Alagon, A
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Pharmacology & Pharmacy
Toxicology
Loxosceles
recombinant sphingomyelinase D
enzymatic activity
antivenom
topic Pharmacology & Pharmacy
Toxicology
Loxosceles
recombinant sphingomyelinase D
enzymatic activity
antivenom
description We report the cloning of sphingomyelinase D (SMD) cDNA from Loxosceles reclusa, Loxosceles boneti and Loxosceles laeta into bacterial expression systems, as well as optimization of expression conditions so as to obtain soluble and active recombinant enzymes. The recombinant mature SMDs, tagged with a histidine tail at the N- or C-termini, were compared in terms of toxicity and enzymatic activity, and were used as immunogens for the production of monovalent antiscra in rabbits and F(ab')(2) preparations in animals used for commercial antivenom production (horses). We performed studies on in vitro inhibition of enzymatic activity of natural venom preparations by antibodies generated against the tagged proteins. We also present and discuss the results of studies on the specific and para-specific in vivo protective potential of the rabbit and equine antibody preparations against the recombinant proteins themselves and natural venom preparations. Our conclusions support the feasibility of using recombinant SMDs for production and evaluation of polyvalent anti-Loxosceles antivenoms, and we offer data on the potential of paraspecific neutralization in the context of the antigenic groupings and the molecular phylogeny of those active SMDs for which amino acid sequence information is available. (c) 2006 Elsevier Ltd. All rights reserved.
publishDate 2006
dc.date.none.fl_str_mv 2006
2011-01-22T10:26:24Z
2011-01-22T10:26:24Z
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv 0041-0101
http://hdl.handle.net/11154/3190
1348
10.1016/j.toxicon.2006.04.010
identifier_str_mv 0041-0101
1348
10.1016/j.toxicon.2006.04.010
url http://hdl.handle.net/11154/3190
dc.language.none.fl_str_mv en
language_invalid_str_mv en
dc.relation.none.fl_str_mv Toxicon
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.source.none.fl_str_mv 48(1):64-74
reponame:Sistema de Información de la Facultad de Ciencias, UNAM
instname:Universidad Nacional Autónoma de México
instacron:UNAM
instname_str Universidad Nacional Autónoma de México
instacron_str UNAM
institution UNAM
reponame_str Sistema de Información de la Facultad de Ciencias, UNAM
collection Sistema de Información de la Facultad de Ciencias, UNAM
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1858175401757507584
score 15,811543

Similares en LA Referencia