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

hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorVALLET-COURBIN, Amelie
dc.contributor.authorLARIVIERE, Melusine
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorHOCQUELLET, Agnes
dc.contributor.authorHEMADOU, Audrey
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorPARIMALA, Sarjapura-Nagaraja
dc.contributor.authorLAROCHE-TRAINEAU, Jeanny
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorSANTARELLI, Xavier
dc.contributor.authorCLOFENT-SANCHEZ, Gisele
dc.contributor.authorJACOBIN-VALAT, Marie-Josee
hal.structure.identifierChimie et Biologie des Membranes et des Nanoobjets [CBMN]
dc.contributor.authorNOUBHANI, Abdelmajid
dc.date.accessioned2020-07-09T14:16:43Z
dc.date.available2020-07-09T14:16:43Z
dc.date.issued2017
dc.identifier.urihttps://oskar-bordeaux.fr/handle/20.500.12278/10290
dc.description.abstractEnCells of the innate and adaptive immune system are key factors in the progression of atherosclerotic plaque, leading to plaque instability and rupture, potentially resulting in acute atherothrombotic 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-alphaIIbbeta3 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-alphaIIbbeta3 TEG4 scFv previously obtained from a phage-display selection on activated platelets, was inserted into the eukaryote vector (pPICZalphaA) 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.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.halChimie/Matériaux
bordeaux.journalPloS one
bordeaux.pagee0170305-e0170305
bordeaux.volume12
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN) - UMR 5248*
bordeaux.hal.laboratoriesInstitut de Chimie & de Biologie des Membranes & des Nano-objets (CBMN, UMR 5248)
bordeaux.issue1
bordeaux.institutionUniversité de Bordeaux
bordeaux.institutionBordeaux INP
bordeaux.COinSctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.jtitle=PloS%20one&rft.date=2017&rft.volume=12&rft.issue=1&rft.spage=e0170305-e0170305&rft.epage=e0170305-e0170305&rft.au=VALLET-COURBIN,%20Amelie&LARIVIERE,%20Melusine&HOCQUELLET,%20Agnes&HEMADOU,%20Audrey&PARIMALA,%20Sarjapura-Nagaraja&rft.genre=article


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