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

dc.creatorFernandes, Cláudia Pinho Hartleben
dc.creatorSeixas, Fabiana K
dc.creatorCoutinho, Mariana Loner
dc.creatorVasconcellos, Flávia A
dc.creatorSeyffert, Núbia
dc.creatorCroda, Julio Henrique Rosa
dc.creatorMcbride, Alan John Alexander
dc.creatorKo, Albert Icksang
dc.creatorDellagostin, Odir Antonio
dc.creatorAleixo, José Antonio Guimarães
dc.date2015-11-10T14:00:35Z
dc.date2015-11-10T14:00:35Z
dc.date2007
dc.date.accessioned2023-09-26T20:27:34Z
dc.date.available2023-09-26T20:27:34Z
dc.identifierFERNANDES, 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.identifier1554-0014
dc.identifier10.1089/hyb.2006.033
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/12144
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8857380
dc.descriptionPathogenic 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.
dc.formatapplication/pdf
dc.languageeng
dc.publisherMary Ann Lieberty
dc.rightsopen access
dc.subjectAnticorpos Monoclonais/fisiologia
dc.subjectProteínas da Membrana Bacteriana Externa/imunologia
dc.subjectLeptospira/imunologia
dc.subjectLeptospirose/diagnóstico
dc.subjectLipoproteínas/imunologia
dc.subjectAnimais
dc.subjectAnticorpos Monoclonais/biossíntese
dc.subjectAnticorpos Monoclonais/uso diagnóstico
dc.subjectFeminino
dc.subjectLeptospira/patogenicidade
dc.subjectLeptospirose/microbiologia
dc.subjectCamundongos
dc.subjectCamundongos Endogâmicos BALB C
dc.titleMonoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
dc.typeArticle


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