Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)

Detalhes bibliográficos
Autor(a) principal: Scharfstein, Julio
Data de Publicação: 1995
Outros Autores: Abrahamson, Magnus, Souza, Clarissa Beatriz Palatnik de, Barral, Aldina Maria Prado, Silva, Ian Victor
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UFBA
Texto Completo: http://www.repositorio.ufba.br/ri/handle/ri/8543
Resumo: Texto completo: acesso restrito. p.63–72
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spelling Scharfstein, JulioAbrahamson, MagnusSouza, Clarissa Beatriz Palatnik deBarral, Aldina Maria PradoSilva, Ian VictorScharfstein, JulioAbrahamson, MagnusSouza, Clarissa Beatriz Palatnik deBarral, Aldina Maria PradoSilva, Ian Victor2013-02-19T16:29:55Z19950022-1759http://www.repositorio.ufba.br/ri/handle/ri/8543v. 182, n. 1Texto completo: acesso restrito. p.63–72A novel immunoassay (PINC-ELISA) was designed using proteinase inhibitors of the cystatin superfamily (PINC) in the solid phase, to promote the selective capture of cysteine proteinases. The method was applied in the identification of papain-like antigens from parasitic protozoa. PINC of human origin, namely recombinant cystatin C (r-cystatin C) or low molecular weight kininogen were used in the assays to adsorb proteases contained in cell lysates from various trypanosomatids. The PINC-ELISA was at first optimized with the major cysteine proteinase from Trypanosoma cruzi (known as View the MathML source or cruzipain), an antigen whose serodiagnostic properties were previously established. Cruzipain is selectively adsorbed from crude extracts of T. cruzi onto PINC-coated wells; the finding that antibodies bind to epitopes located away from the sites of interaction with r-cystatin or low molecular weight kininogen has allowed for the screening of antibodies in chagasic sera, the methodology being advantageous in that it dispensed prior purification of the proteinase antigen. The PINC-ELISA was then carried out with lysates originating from Leishmania m. amazonensis (amastigotes) or Leishmania donovani (promastigotes). Complexes between solid-phase r-cystatin C and antigenic ligands in the lysates were again detected. The Leishmania molecules which bound to r-cystatin C, were respectively recognized by serum antibodies from mice chronically infected with L. amazonensis or from patients with visceral leishmaniasis. Direct evidence for the presence of cysteine proteinases in lysates from L. donovani was then obtained, using synthetic fluorogenic substrates. Due to the broad inhibition profile of r-cystatin C and the marked antigenicity of parasitic cysteine proteinases, such enzymes can be readily detected by PINC-ELISA, without requirement for prior knowledge of their substrate specificities.Submitted by Suelen Reis (suelen_suzane@hotmail.com) on 2013-02-18T16:42:04Z No. of bitstreams: 1 Scharfstein.pdf: 999028 bytes, checksum: 0517c0b32a8bbb053e47114885149a4d (MD5)Approved for entry into archive by Fatima Cleômenis Botelho Maria (botelho@ufba.br) on 2013-02-19T16:29:54Z (GMT) No. of bitstreams: 1 Scharfstein.pdf: 999028 bytes, checksum: 0517c0b32a8bbb053e47114885149a4d (MD5)Made available in DSpace on 2013-02-19T16:29:55Z (GMT). No. of bitstreams: 1 Scharfstein.pdf: 999028 bytes, checksum: 0517c0b32a8bbb053e47114885149a4d (MD5) Previous issue date: 1995http://dx.doi.org/10.1016/0022-1759(95)00023-4reponame:Repositório Institucional da UFBAinstname:Universidade Federal da Bahia (UFBA)instacron:UFBACysteine proteinaseCystatinKininogenImmunoassayTrypanosoma cruziLeishmaniaAntigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)Journal of Immunological Methodsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article10000-01-01enginfo:eu-repo/semantics/openAccessORIGINALScharfstein.pdfScharfstein.pdfapplication/pdf999028https://repositorio.ufba.br/bitstream/ri/8543/1/Scharfstein.pdf0517c0b32a8bbb053e47114885149a4dMD51LICENSElicense.txtlicense.txttext/plain1762https://repositorio.ufba.br/bitstream/ri/8543/2/license.txt1b89a9a0548218172d7c829f87a0eab9MD52TEXTScharfstein.pdf.txtScharfstein.pdf.txtExtracted texttext/plain36678https://repositorio.ufba.br/bitstream/ri/8543/3/Scharfstein.pdf.txt936174a0f200ce73de9f1ee3af12e1ebMD53ri/85432022-07-05 14:03:47.789oai:repositorio.ufba.br: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Repositório InstitucionalPUBhttp://192.188.11.11:8080/oai/requestopendoar:19322022-07-05T17:03:47Repositório Institucional da UFBA - Universidade Federal da Bahia (UFBA)false
dc.title.pt_BR.fl_str_mv Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
dc.title.alternative.pt_BR.fl_str_mv Journal of Immunological Methods
title Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
spellingShingle Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
Scharfstein, Julio
Cysteine proteinase
Cystatin
Kininogen
Immunoassay
Trypanosoma cruzi
Leishmania
title_short Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
title_full Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
title_fullStr Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
title_full_unstemmed Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
title_sort Antigenicity of cystatin-binding proteins from parasitic protozoan Detection by a proteinase inhibitor based capture immunoassay (PINC-ELISA)
author Scharfstein, Julio
author_facet Scharfstein, Julio
Abrahamson, Magnus
Souza, Clarissa Beatriz Palatnik de
Barral, Aldina Maria Prado
Silva, Ian Victor
author_role author
author2 Abrahamson, Magnus
Souza, Clarissa Beatriz Palatnik de
Barral, Aldina Maria Prado
Silva, Ian Victor
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Scharfstein, Julio
Abrahamson, Magnus
Souza, Clarissa Beatriz Palatnik de
Barral, Aldina Maria Prado
Silva, Ian Victor
Scharfstein, Julio
Abrahamson, Magnus
Souza, Clarissa Beatriz Palatnik de
Barral, Aldina Maria Prado
Silva, Ian Victor
dc.subject.por.fl_str_mv Cysteine proteinase
Cystatin
Kininogen
Immunoassay
Trypanosoma cruzi
Leishmania
topic Cysteine proteinase
Cystatin
Kininogen
Immunoassay
Trypanosoma cruzi
Leishmania
description Texto completo: acesso restrito. p.63–72
publishDate 1995
dc.date.issued.fl_str_mv 1995
dc.date.accessioned.fl_str_mv 2013-02-19T16:29:55Z
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.uri.fl_str_mv http://www.repositorio.ufba.br/ri/handle/ri/8543
dc.identifier.issn.none.fl_str_mv 0022-1759
dc.identifier.number.pt_BR.fl_str_mv v. 182, n. 1
identifier_str_mv 0022-1759
v. 182, n. 1
url http://www.repositorio.ufba.br/ri/handle/ri/8543
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.source.pt_BR.fl_str_mv http://dx.doi.org/10.1016/0022-1759(95)00023-4
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFBA
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