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A Recombinant Human Anti-Platelet scFv Antibody Produced in Pichia pastoris for Atheroma Targeting

hal.structure.identifierInstitut des Sciences de la Vigne et du Vin (ISVV)
dc.contributor.authorVALLET-COURBIN, Amélie
dc.contributor.authorLARIVIÈRE, Mélusine
hal.structure.identifierBiotechnologie des protéines recombinantes à visée santé
dc.contributor.authorHOCQUELLET, Agnès
hal.structure.identifierCentre de résonance magnétique des systèmes biologiques [CRMSB]
dc.contributor.authorHEMADOU, Audrey
dc.contributor.authorPARIMALA, Sarjapura-Nagaraja
dc.contributor.authorLAROCHE-TRAINEAU, Jeanny
hal.structure.identifierUniversité Sciences et Technologies - Bordeaux 1 [UB]
dc.contributor.authorSANTARELLI, Xavier
dc.contributor.authorCLOFENT-SANCHEZ, Gisèle
dc.contributor.authorJACOBIN-VALAT, Marie-Josée
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorNOUBHANI, Abdelmajid
dc.date.accessioned2021-10-07T16:29:32Z
dc.date.available2021-10-07T16:29:32Z
dc.date.issued2017-01-26
dc.identifier.issn1932-6203
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/112657
dc.description.abstractEnCells of the innate and adaptive immune system are key factors in the progression of athero-sclerotic plaque, leading to plaque instability and rupture, potentially resulting in acute ather-othrombotic events such as coronary artery disease, cerebrovascular disease and peripheral arterial disease. Here, we describe the cloning, expression, purification, and immunoreactivity assessment of a recombinant single-chain variable fragment (scFv) derived from a human anti-αIIbβ3 antibody (HuAb) selected to target atheromatous lesions for the presence of platelets. Indeed, platelets within atheroma plaques have been shown to play a role in inflammation, in platelet-leucocyte aggregates and in thrombi formation and might thus be considered relevant biomarkers of atherosclerotic progression. The DNA sequence that encodes the anti-αIIbβ3 TEG4 scFv previously obtained from a phage-display selection on activated platelets, was inserted into the eukaryote vector (pPICZαA) in fusion with a tag sequence encoding 2 cysteines useable for specific probes grafting experiments. The recombinant protein was expressed at high yields in Pichia pastoris (30 mg/L culture). The advantage of P. pastoris as an expression system is the production and secretion of recombinant proteins in the supernatant, ruling out the difficulties encountered when scFv are produced in the cytoplasm of bacteria (low yield, low solubility and reduced affinity). The improved conditions allowed for the recovery of highly purified and biologically active scFv fragments ready to be grafted in a site-directed way to nanoparticles for the imaging of atherosclerotic plaques involving inflammatory processes and thus at high risk of instability.
dc.language.isoen
dc.publisherPublic Library of Science
dc.title.enA Recombinant Human Anti-Platelet scFv Antibody Produced in Pichia pastoris for Atheroma Targeting
dc.typeArticle de revue
dc.identifier.doi10.1371/journal.pone.0170305
dc.subject.halSciences du Vivant [q-bio]/Immunologie
bordeaux.journalPLoS ONE
bordeaux.pagee0170305
bordeaux.volume12
bordeaux.hal.laboratoriesCentre de Résonance Magnétique des Systèmes Biologiques (CRMSB) - UMR 5536*
bordeaux.issue1
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionCNRS
bordeaux.peerReviewedoui
hal.identifierhal-02376377
hal.version1
hal.origin.linkhttps://hal.archives-ouvertes.fr//hal-02376377v1
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=PLoS%20ONE&rft.date=2017-01-26&rft.volume=12&rft.issue=1&rft.spage=e0170305&rft.epage=e0170305&rft.eissn=1932-6203&rft.issn=1932-6203&rft.au=VALLET-COURBIN,%20Am%C3%A9lie&LARIVI%C3%88RE,%20M%C3%A9lusine&HOCQUELLET,%20Agn%C3%A8s&HEMADOU,%20Audrey&PARIMALA,%20Sarjapura-Nagaraja&rft.genre=article


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