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

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dc.contributor.authorSemedo, Madga Sofia Cardoso Nobre
dc.contributor.authorKarmali, Amin
dc.contributor.authorMartins, Sónia
dc.contributor.authorFonseca, Luís
dc.date.accessioned2016-04-28T11:03:20Z
dc.date.available2016-04-28T11:03:20Z
dc.date.issued2016
dc.description.abstractβ-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.pt_PT
dc.identifier.citationSEMEDO, 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. 333-342pt_PT
dc.identifier.doi10.1016/j.procbio.2015.11.029pt_PT
dc.identifier.issn1359-5113
dc.identifier.urihttp://hdl.handle.net/10400.21/6114
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherElsevier Ltdpt_PT
dc.subjectBasidiomycete mushroomspt_PT
dc.subjectEpitope analysis of Mabspt_PT
dc.subjectExtracellular -d-glucans from Pleurotuspt_PT
dc.subjectHybridoma technologypt_PT
dc.subjectMabs of IgG classpt_PT
dc.subjectOstreatuspt_PT
dc.subjectSpecificity and cross-reactivitypt_PT
dc.titleGeneration of high-affinity monoclonal antibodies of IgG class against native β-d-glucans from basidiomycete mushroomspt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage342pt_PT
oaire.citation.issue2pt_PT
oaire.citation.startPage333pt_PT
oaire.citation.titleProcess Biochemistrypt_PT
oaire.citation.volume51pt_PT
person.familyNameSemedo
person.familyNameKarmali
person.givenNameMagda
person.givenNameAmin
person.identifier1022083
person.identifier.ciencia-id9C11-BD02-C77F
person.identifier.orcid0000-0001-6330-2899
person.identifier.orcid0000-0003-0419-401X
person.identifier.ridE-4787-2011
person.identifier.scopus-author-id57000381900
person.identifier.scopus-author-id6603778216
rcaap.rightsclosedAccesspt_PT
rcaap.typearticlept_PT
relation.isAuthorOfPublication73e398bc-a0e8-420d-81eb-34159398838d
relation.isAuthorOfPublication23975b4b-3234-4139-9675-89ea6f9a2332
relation.isAuthorOfPublication.latestForDiscovery23975b4b-3234-4139-9675-89ea6f9a2332

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