Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.

Detalhes bibliográficos
Autor(a) principal: Fernandes, Cláudia Pinho Hartleben
Data de Publicação: 2007
Outros Autores: Seixas, Fabiana K, Coutinho, Mariana Loner, Vasconcellos, Flávia A, Seyffert, Núbia, Croda, Julio Henrique Rosa, Mcbride, Alan John Alexander, Ko, Albert Icksang, Dellagostin, Odir Antonio, Aleixo, José Antonio Guimarães
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/12144
Resumo: Centro de Biotecnologia. Pelotas, RS, Brasil / Universidade Federal de Pelotas. Faculdade de Medicina Veterinária. Pelotas, RS, Brasil
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spelling Fernandes, Cláudia Pinho HartlebenSeixas, Fabiana KCoutinho, Mariana LonerVasconcellos, Flávia ASeyffert, NúbiaCroda, Julio Henrique RosaMcbride, Alan John AlexanderKo, Albert IcksangDellagostin, Odir AntonioAleixo, José Antonio Guimarães2015-11-10T14:00:35Z2015-11-10T14:00:35Z2007FERNANDES, C. P. H. et al. Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications. Hybridoma (Larchmt), v. 26, n. 1, p. 35-41, 2007.1554-001410.1089/hyb.2006.033https://www.arca.fiocruz.br/handle/icict/12144engMary Ann LiebertyMonoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleCentro de Biotecnologia. Pelotas, RS, Brasil / Universidade Federal de Pelotas. Faculdade de Medicina Veterinária. Pelotas, RS, BrasilCentro de Biotecnologia. Pelotas, RS, BrasilCentro de Biotecnologia. Pelotas, RS, BrasilCentro de Biotecnologia. Pelotas, RS, BrasilCentro de Biotecnologia. Pelotas, RS, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Cornell University. Weill Medical College. Division of International Medicine and Infectious Disease. New YorkCentro de Biotecnologia. Pelotas, RS, BrasilCentro de Biotecnologia. Pelotas, RS, BrasilPathogenic serovars of Leptospira have a wide antigenic diversity attributed mainly to the lipopolysaccharide present in the outer membrane. In contrast, antigens conserved among pathogenic serovars are mainly represented by outer membrane proteins. Surface exposure of a major and highly conserved outer membrane lipoprotein (LipL32) was recently demonstrated on pathogenic Leptospira. LipL32 in its recombinant form (rLipL32) was used to immunize BALB/c mice to develop murine monoclonal antibodies (MAbs). Three MAbs against rLipL32 were produced, isotyped, and evaluated for further use in diagnostic tests of leptospirosis using different approaches. MAbs were conjugated to peroxidase and evaluated in a native protein enzyme-linked immunosorbent assay (ELISA) with intact and heat-treated leptospiral cells, conjugated to fluorescein isothiocyanate (FITC) for indirect immunofluorescence with intact and methanol fixed cells and were used for LipL32 immunoprecipitation from leptospiral cells. rLipL32 MAbs conjugated to peroxidase or used as primary antibody bound to intact and heat-treated cells in ELISA, proving that they could be used in enzyme immunoassays for detection of the native protein. In immunofluorescence assay, MAbs labeled bacterial cells either intact or methanol fixed. Two MAbs were able to immunoprecipitate the native protein from live and motile leptospiral cells and, adsorbed onto magnetic beads, captured intact bacteria from artificially contaminated human sera for detection by polymerase chain reaction (PCR) amplification. Results of this study suggest that the MAbs produced can be useful for the development of diagnostic tests based on detection of LipL32 leptospiral antigen in biological fluids.Anticorpos Monoclonais/fisiologiaProteínas da Membrana Bacteriana Externa/imunologiaLeptospira/imunologiaLeptospirose/diagnósticoLipoproteínas/imunologiaAnimaisAnticorpos Monoclonais/biossínteseAnticorpos Monoclonais/uso diagnósticoFemininoLeptospira/patogenicidadeLeptospirose/microbiologiaCamundongosCamundongos Endogâmicos BALB Cinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82354https://www.arca.fiocruz.br/bitstream/icict/12144/1/license.txt8b4c200b4e10021c5683c6ccaba07169MD51ORIGINALFernandes CPH Monoclonal antibodies against....pdfFernandes CPH Monoclonal antibodies against....pdfapplication/pdf2615824https://www.arca.fiocruz.br/bitstream/icict/12144/2/Fernandes%20CPH%20Monoclonal%20antibodies%20against....pdf6323db4939230d4e82e18d306c86946cMD52TEXTFernandes CPH Monoclonal antibodies against....pdf.txtFernandes CPH Monoclonal antibodies against....pdf.txtExtracted texttext/plain31279https://www.arca.fiocruz.br/bitstream/icict/12144/3/Fernandes%20CPH%20Monoclonal%20antibodies%20against....pdf.txtc62489fbf401384cce4e794958ecd130MD53icict/121442023-03-15 14:33:21.758oai:www.arca.fiocruz.br:icict/12144Q0VTU8ODTyBOw4NPLUVYQ0xVU0lWQSBERSBESVJFSVRPUyBBVVRPUkFJUwoKQW8gYWNlaXRhciBvcyBURVJNT1MgZSBDT05EScOHw5VFUyBkZXN0YSBDRVNTw4NPLCBvIEFVVE9SIGUvb3UgVElUVUxBUiBkZSBkaXJlaXRvcyBhdXRvcmFpcyBzb2JyZSBhIE9CUkEgZGUgcXVlIHRyYXRhIGVzdGUgZG9jdW1lbnRvOgoKQ0VERSBlIFRSQU5TRkVSRSwgdG90YWwgZSBncmF0dWl0YW1lbnRlLCDDoCBGSU9DUlVaIC0gRlVOREHDh8ODTyBPU1dBTERPIENSVVosIGVtIGNhcsOhdGVyIHBlcm1hbmVudGUsIGlycmV2b2fDoXZlbCBlIE7Dg08gRVhDTFVTSVZPLCAKdG9kb3Mgb3MgZGlyZWl0b3MgcGF0cmltb25pYWlzIE7Dg08gQ09NRVJDSUFJUyBkZSB1dGlsaXphw6fDo28gZGEgT0JSQSBhcnTDrXN0aWNhIGUvb3UgY2llbnTDrWZpY2EgaW5kaWNhZGEgYWNpbWEsIGluY2x1c2l2ZSBvcyBkaXJlaXRvcyAKZGUgdm96IGUgaW1hZ2VtIHZpbmN1bGFkb3Mgw6AgT0JSQSwgZHVyYW50ZSB0b2RvIG8gcHJhem8gZGUgZHVyYcOnw6NvIGRvcyBkaXJlaXRvcyBhdXRvcmFpcywgZW0gcXVhbHF1ZXIgaWRpb21hIGUgZW0gdG9kb3Mgb3MgcGHDrXNlczsKICAgICAgICAKQUNFSVRBIHF1ZSBhIGNlc3PDo28gdG90YWwgbsOjbyBleGNsdXNpdmEsIHBlcm1hbmVudGUgZSBpcnJldm9nw6F2ZWwgZG9zIGRpcmVpdG9zIGF1dG9yYWlzIHBhdHJpbW9uaWFpcyBuw6NvIGNvbWVyY2lhaXMgZGUgdXRpbGl6YcOnw6NvIApkZSBxdWUgdHJhdGEgZXN0ZSBkb2N1bWVudG8gaW5jbHVpLCBleGVtcGxpZmljYXRpdmFtZW50ZSwgb3MgZGlyZWl0b3MgZGUgZGlzcG9uaWJpbGl6YcOnw6NvIGUgY29tdW5pY2HDp8OjbyBww7pibGljYSBkYSBPQlJBLCBlbSBxdWFscXVlciAKbWVpbyBvdSB2ZcOtY3VsbywgaW5jbHVzaXZlIGVtIFJlcG9zaXTDs3Jpb3MgRGlnaXRhaXMsIGJlbSBjb21vIG9zIGRpcmVpdG9zIGRlIHJlcHJvZHXDp8OjbywgZXhpYmnDp8OjbywgZXhlY3XDp8OjbywgZGVjbGFtYcOnw6NvLCBleHBvc2nDp8OjbywgCmFycXVpdmFtZW50bywgaW5jbHVzw6NvIGVtIGJhbmNvIGRlIGRhZG9zLCBwcmVzZXJ2YcOnw6NvLCBkaWZ1c8OjbywgZGlzdHJpYnVpw6fDo28sIGRpdnVsZ2HDp8OjbywgZW1wcsOpc3RpbW8sIHRyYWR1w6fDo28sIGluY2x1c8OjbyBlbSBub3ZhcyAKb2JyYXMgb3UgY29sZXTDom5lYXMsIHJldXRpbGl6YcOnw6NvLCBlZGnDp8OjbywgcHJvZHXDp8OjbyBkZSBtYXRlcmlhbCBkaWTDoXRpY28gZSBjdXJzb3Mgb3UgcXVhbHF1ZXIgZm9ybWEgZGUgdXRpbGl6YcOnw6NvIG7Do28gY29tZXJjaWFsOwogICAgICAgIApSRUNPTkhFQ0UgcXVlIGEgY2Vzc8OjbyBhcXVpIGVzcGVjaWZpY2FkYSBjb25jZWRlIMOgIEZJT0NSVVogLSBGVU5EQcOHw4NPIE9TV0FMRE8gQ1JVWiBvIGRpcmVpdG8gZGUgYXV0b3JpemFyIHF1YWxxdWVyIHBlc3NvYSDigJMgZsOtc2ljYSAKb3UganVyw61kaWNhLCBww7pibGljYSBvdSBwcml2YWRhLCBuYWNpb25hbCBvdSBlc3RyYW5nZWlyYSDigJMgYSBhY2Vzc2FyIGUgdXRpbGl6YXIgYW1wbGFtZW50ZSBhIE9CUkEsIHNlbSBleGNsdXNpdmlkYWRlLCBwYXJhIHF1YWlzcXVlciAKZmluYWxpZGFkZXMgbsOjbyBjb21lcmNpYWlzOwogICAgICAgIApERUNMQVJBIHF1ZSBhIG9icmEgw6kgY3JpYcOnw6NvIG9yaWdpbmFsIGUgcXVlIMOpIG8gdGl0dWxhciBkb3MgZGlyZWl0b3MgYXF1aSBjZWRpZG9zIGUgYXV0b3JpemFkb3MsIHJlc3BvbnNhYmlsaXphbmRvLXNlIGludGVncmFsbWVudGUgCnBlbG8gY29udGXDumRvIGUgb3V0cm9zIGVsZW1lbnRvcyBxdWUgZmF6ZW0gcGFydGUgZGEgT0JSQSwgaW5jbHVzaXZlIG9zIGRpcmVpdG9zIGRlIHZveiBlIGltYWdlbSB2aW5jdWxhZG9zIMOgIE9CUkEsIG9icmlnYW5kby1zZSBhIAppbmRlbml6YXIgdGVyY2Vpcm9zIHBvciBkYW5vcywgYmVtIGNvbW8gaW5kZW5pemFyIGUgcmVzc2FyY2lyIMOgIEZJT0NSVVogLSBGVU5EQcOHw4NPIE9TV0FMRE8gQ1JVWiBkZSBldmVudHVhaXMgZGVzcGVzYXMgcXVlIHZpZXJlbSBhIApzdXBvcnRhciwgZW0gcmF6w6NvIGRlIHF1YWxxdWVyIG9mZW5zYSBhIGRpcmVpdG9zIGF1dG9yYWlzIG91IGRpcmVpdG9zIGRlIHZveiBvdSBpbWFnZW0sIHByaW5jaXBhbG1lbnRlIG5vIHF1ZSBkaXogcmVzcGVpdG8gYSBwbMOhZ2lvIAplIHZpb2xhw6fDtWVzIGRlIGRpcmVpdG9zOwogICAgICAgIApBRklSTUEgcXVlIGNvbmhlY2UgYSBQb2zDrXRpY2EgSW5zdGl0dWNpb25hbCBkZSBBY2Vzc28gQWJlcnRvIGRhIEluc3RpdHVpw6fDo28gZSBhcyBkaXJldHJpemVzIHBhcmEgbyBmdW5jaW9uYW1lbnRvIGRvIHJlcG9zaXTDs3JpbyAKaW5zdGl0dWNpb25hbCBBUkNBLgo=Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352023-03-15T17:33:21Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
title Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
spellingShingle Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
Fernandes, Cláudia Pinho Hartleben
Anticorpos Monoclonais/fisiologia
Proteínas da Membrana Bacteriana Externa/imunologia
Leptospira/imunologia
Leptospirose/diagnóstico
Lipoproteínas/imunologia
Animais
Anticorpos Monoclonais/biossíntese
Anticorpos Monoclonais/uso diagnóstico
Feminino
Leptospira/patogenicidade
Leptospirose/microbiologia
Camundongos
Camundongos Endogâmicos BALB C
title_short Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
title_full Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
title_fullStr Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
title_full_unstemmed Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
title_sort Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
author Fernandes, Cláudia Pinho Hartleben
author_facet Fernandes, Cláudia Pinho Hartleben
Seixas, Fabiana K
Coutinho, Mariana Loner
Vasconcellos, Flávia A
Seyffert, Núbia
Croda, Julio Henrique Rosa
Mcbride, Alan John Alexander
Ko, Albert Icksang
Dellagostin, Odir Antonio
Aleixo, José Antonio Guimarães
author_role author
author2 Seixas, Fabiana K
Coutinho, Mariana Loner
Vasconcellos, Flávia A
Seyffert, Núbia
Croda, Julio Henrique Rosa
Mcbride, Alan John Alexander
Ko, Albert Icksang
Dellagostin, Odir Antonio
Aleixo, José Antonio Guimarães
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Fernandes, Cláudia Pinho Hartleben
Seixas, Fabiana K
Coutinho, Mariana Loner
Vasconcellos, Flávia A
Seyffert, Núbia
Croda, Julio Henrique Rosa
Mcbride, Alan John Alexander
Ko, Albert Icksang
Dellagostin, Odir Antonio
Aleixo, José Antonio Guimarães
dc.subject.decs.pt_BR.fl_str_mv Anticorpos Monoclonais/fisiologia
Proteínas da Membrana Bacteriana Externa/imunologia
Leptospira/imunologia
Leptospirose/diagnóstico
Lipoproteínas/imunologia
Animais
Anticorpos Monoclonais/biossíntese
Anticorpos Monoclonais/uso diagnóstico
Feminino
Leptospira/patogenicidade
Leptospirose/microbiologia
Camundongos
Camundongos Endogâmicos BALB C
topic Anticorpos Monoclonais/fisiologia
Proteínas da Membrana Bacteriana Externa/imunologia
Leptospira/imunologia
Leptospirose/diagnóstico
Lipoproteínas/imunologia
Animais
Anticorpos Monoclonais/biossíntese
Anticorpos Monoclonais/uso diagnóstico
Feminino
Leptospira/patogenicidade
Leptospirose/microbiologia
Camundongos
Camundongos Endogâmicos BALB C
description Centro de Biotecnologia. Pelotas, RS, Brasil / Universidade Federal de Pelotas. Faculdade de Medicina Veterinária. Pelotas, RS, Brasil
publishDate 2007
dc.date.issued.fl_str_mv 2007
dc.date.accessioned.fl_str_mv 2015-11-10T14:00:35Z
dc.date.available.fl_str_mv 2015-11-10T14:00:35Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv FERNANDES, C. P. H. et al. Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications. Hybridoma (Larchmt), v. 26, n. 1, p. 35-41, 2007.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/12144
dc.identifier.issn.none.fl_str_mv 1554-0014
10.1089/hyb.2006.033
identifier_str_mv FERNANDES, C. P. H. et al. Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications. Hybridoma (Larchmt), v. 26, n. 1, p. 35-41, 2007.
1554-0014
10.1089/hyb.2006.033
url https://www.arca.fiocruz.br/handle/icict/12144
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Mary Ann Lieberty
publisher.none.fl_str_mv Mary Ann Lieberty
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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https://www.arca.fiocruz.br/bitstream/icict/12144/2/Fernandes%20CPH%20Monoclonal%20antibodies%20against....pdf
https://www.arca.fiocruz.br/bitstream/icict/12144/3/Fernandes%20CPH%20Monoclonal%20antibodies%20against....pdf.txt
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