Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms

Detalhes bibliográficos
Autor(a) principal: Semedo, Madga Sofia Cardoso Nobre
Data de Publicação: 2016
Outros Autores: Karmali, Amin, Martins, Sónia, Fonseca, Luís
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.21/6114
Resumo: β-d-glucans from basidiomycete strains are powerful immunomodulatory agents in several clinical conditions. Therefore, their assay, purification and characterization are of great interest to understand their structure-function relationship. Hybridoma cell fusion was used to raise monoclonal antibodies (Mabs) against extracellular β-d-glucans (EBGs) from Pleurotus ostreatus. Two of the hybridoma clones (1E6-1E8-B5 and 3E8-3B4) secreting Mabs against EBGs were selected. This hybridoma cell line secreted Mabs of the IgG class which were then purified by hydroxyapatite chromatography to apparent homogeneity on native and SDS-PAGE. Mabs secreted by 1E6-1E8-B5 clone were found to recognize a common epitope on several β-d-glucans from different basidiomycete strains. This Mab exhibited high affinity constant (KA) for β-d-glucans from several mushroom strains in the range of 3.20 × 109 ± 3.32 × 103-1.51 × 1013 ± 3.58 × 107 L/mol. Moreover, they reacted to some heat-treated β-d-glucans in a different mode when compared with the native forms; these data suggest that this Mab binds to a conformational epitope on the β-d-glucan molecule. The epitope-binding studies of Mabs obtained from 1E6-1E8-B5 and 3E8-3B4 revealed that the Mabs bind to the same epitope on some β-d-glucans and to different epitopes in other antigen molecules. Therefore, these Mabs can be used to assay for β-d-glucan from basidiomycete mushrooms. © 2015 Elsevier Ltd. All rights reserved.
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spelling Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushroomsBasidiomycete mushroomsEpitope analysis of MabsExtracellular -d-glucans from PleurotusHybridoma technologyMabs of IgG classOstreatusSpecificity and cross-reactivityβ-d-glucans from basidiomycete strains are powerful immunomodulatory agents in several clinical conditions. Therefore, their assay, purification and characterization are of great interest to understand their structure-function relationship. Hybridoma cell fusion was used to raise monoclonal antibodies (Mabs) against extracellular β-d-glucans (EBGs) from Pleurotus ostreatus. Two of the hybridoma clones (1E6-1E8-B5 and 3E8-3B4) secreting Mabs against EBGs were selected. This hybridoma cell line secreted Mabs of the IgG class which were then purified by hydroxyapatite chromatography to apparent homogeneity on native and SDS-PAGE. Mabs secreted by 1E6-1E8-B5 clone were found to recognize a common epitope on several β-d-glucans from different basidiomycete strains. This Mab exhibited high affinity constant (KA) for β-d-glucans from several mushroom strains in the range of 3.20 × 109 ± 3.32 × 103-1.51 × 1013 ± 3.58 × 107 L/mol. Moreover, they reacted to some heat-treated β-d-glucans in a different mode when compared with the native forms; these data suggest that this Mab binds to a conformational epitope on the β-d-glucan molecule. The epitope-binding studies of Mabs obtained from 1E6-1E8-B5 and 3E8-3B4 revealed that the Mabs bind to the same epitope on some β-d-glucans and to different epitopes in other antigen molecules. Therefore, these Mabs can be used to assay for β-d-glucan from basidiomycete mushrooms. © 2015 Elsevier Ltd. All rights reserved.Elsevier LtdRCIPLSemedo, Madga Sofia Cardoso NobreKarmali, AminMartins, SóniaFonseca, Luís2016-04-28T11:03:20Z20162016-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.21/6114engSEMEDO, Magda; [et al] - Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms. Process Biochemistry. ISSN 1359-5113. Vol. 51, N.º 2 (2016), pp. 333-3421359-511310.1016/j.procbio.2015.11.029metadata only accessinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-08-03T09:50:30Zoai:repositorio.ipl.pt:10400.21/6114Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:15:18.834817Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
title Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
spellingShingle Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
Semedo, Madga Sofia Cardoso Nobre
Basidiomycete mushrooms
Epitope analysis of Mabs
Extracellular -d-glucans from Pleurotus
Hybridoma technology
Mabs of IgG class
Ostreatus
Specificity and cross-reactivity
title_short Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
title_full Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
title_fullStr Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
title_full_unstemmed Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
title_sort Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms
author Semedo, Madga Sofia Cardoso Nobre
author_facet Semedo, Madga Sofia Cardoso Nobre
Karmali, Amin
Martins, Sónia
Fonseca, Luís
author_role author
author2 Karmali, Amin
Martins, Sónia
Fonseca, Luís
author2_role author
author
author
dc.contributor.none.fl_str_mv RCIPL
dc.contributor.author.fl_str_mv Semedo, Madga Sofia Cardoso Nobre
Karmali, Amin
Martins, Sónia
Fonseca, Luís
dc.subject.por.fl_str_mv Basidiomycete mushrooms
Epitope analysis of Mabs
Extracellular -d-glucans from Pleurotus
Hybridoma technology
Mabs of IgG class
Ostreatus
Specificity and cross-reactivity
topic Basidiomycete mushrooms
Epitope analysis of Mabs
Extracellular -d-glucans from Pleurotus
Hybridoma technology
Mabs of IgG class
Ostreatus
Specificity and cross-reactivity
description β-d-glucans from basidiomycete strains are powerful immunomodulatory agents in several clinical conditions. Therefore, their assay, purification and characterization are of great interest to understand their structure-function relationship. Hybridoma cell fusion was used to raise monoclonal antibodies (Mabs) against extracellular β-d-glucans (EBGs) from Pleurotus ostreatus. Two of the hybridoma clones (1E6-1E8-B5 and 3E8-3B4) secreting Mabs against EBGs were selected. This hybridoma cell line secreted Mabs of the IgG class which were then purified by hydroxyapatite chromatography to apparent homogeneity on native and SDS-PAGE. Mabs secreted by 1E6-1E8-B5 clone were found to recognize a common epitope on several β-d-glucans from different basidiomycete strains. This Mab exhibited high affinity constant (KA) for β-d-glucans from several mushroom strains in the range of 3.20 × 109 ± 3.32 × 103-1.51 × 1013 ± 3.58 × 107 L/mol. Moreover, they reacted to some heat-treated β-d-glucans in a different mode when compared with the native forms; these data suggest that this Mab binds to a conformational epitope on the β-d-glucan molecule. The epitope-binding studies of Mabs obtained from 1E6-1E8-B5 and 3E8-3B4 revealed that the Mabs bind to the same epitope on some β-d-glucans and to different epitopes in other antigen molecules. Therefore, these Mabs can be used to assay for β-d-glucan from basidiomycete mushrooms. © 2015 Elsevier Ltd. All rights reserved.
publishDate 2016
dc.date.none.fl_str_mv 2016-04-28T11:03:20Z
2016
2016-01-01T00:00:00Z
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://hdl.handle.net/10400.21/6114
url http://hdl.handle.net/10400.21/6114
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv SEMEDO, Magda; [et al] - Generation of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushrooms. Process Biochemistry. ISSN 1359-5113. Vol. 51, N.º 2 (2016), pp. 333-342
1359-5113
10.1016/j.procbio.2015.11.029
dc.rights.driver.fl_str_mv metadata only access
info:eu-repo/semantics/openAccess
rights_invalid_str_mv metadata only access
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier Ltd
publisher.none.fl_str_mv Elsevier Ltd
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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