Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/12144
Title: Monoclonal antibodies against LipL32, the major outer membrane protein of pathogenic Leptospira: production, characterization, and testing in diagnostic applications.
Authors: 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
Affilliation: Centro de Biotecnologia. Pelotas, RS, Brasil / Universidade Federal de Pelotas. Faculdade de Medicina Veterinária. Pelotas, RS, Brasil
Centro de Biotecnologia. Pelotas, RS, Brasil
Centro de Biotecnologia. Pelotas, RS, Brasil
Centro de Biotecnologia. Pelotas, RS, Brasil
Centro de Biotecnologia. Pelotas, RS, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Cornell University. Weill Medical College. Division of International Medicine and Infectious Disease. New York
Centro de Biotecnologia. Pelotas, RS, Brasil
Centro de Biotecnologia. Pelotas, RS, Brasil
Abstract: Pathogenic 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.
DeCS: 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
Issue Date: 2007
Publisher: Mary Ann Lieberty
Citation: 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.
ISSN: 1554-0014
10.1089/hyb.2006.033
Copyright: open access
Appears in Collections:BA - IGM - Artigos de Periódicos

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