Expression and detection strategies for an scFv fragment retaining the same high affinity than Fab and whole antibody: Implications for therapeutic use in prion diseases

Autor: Sosthène Essono, Fatima Dkhissi, Séverine Padiolleau-Lefevre, Jean-Yves Couraud, Coralie Alexandrenne, Anne Wijkhuisen, Didier Boquet, Gilles Clément, Celine A. Blache
Přispěvatelé: Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut de Biologie et de Technologies de Saclay (IBITECS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, INSERM CIC 0802 (INSERM - CHU de Poitiers), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre hospitalier universitaire de Poitiers (CHU Poitiers)-Université de Poitiers, International Space University (ISU), Laboratoire d'Etudes et de Recherches en Immunoanalyses (LERI), Service de Pharmacologie et Immunoanalyse (SPI), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Médicaments et Technologies pour la Santé (MTS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Pari mutuel urbain (.) (PMU), Laboratoire d'Ingénierie des Anticorps pour la Santé (LIAS), Université de Poitiers-Centre hospitalier universitaire de Poitiers (CHU Poitiers)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Rok vydání: 2006
Předmět:
[SDV.BIO]Life Sciences [q-bio]/Biotechnology
medicine.drug_class
Prions
Immunology
Immunoglobulin Variable Region
Context (language use)
Biology
Monoclonal antibody
law.invention
Prion Diseases
03 medical and health sciences
0302 clinical medicine
Antigen
law
medicine
Escherichia coli
Animals
Humans
Avidity
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biochemistry [q-bio.BM]
Molecular Biology
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
0303 health sciences
Expression vector
Antibodies
Monoclonal
[SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Molecular biology
[SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy
Molecular biology
In vitro
Recombinant Proteins
3. Good health
Biochemistry
[SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology
biology.protein
Recombinant DNA
Antibody
030217 neurology & neurosurgery
Zdroj: Molecular Immunology
Molecular Immunology, Elsevier, 2007, 44 (8), pp.1888-1896. ⟨10.1016/j.molimm.2006.09.035⟩
Molecular Immunology, 2007, 44 (8), pp.1888-1896. ⟨10.1016/j.molimm.2006.09.035⟩
ISSN: 0161-5890
DOI: 10.1016/j.molimm.2006.09.035⟩
Popis: Since antibodies currently constitute the most rapidly growing class of human therapeutics, the high-yield production of recombinant antibodies and antibody fragments is a real challenge. Using as model a monoclonal antibody directed against the human prion protein that we prepared previously and tested for its therapeutic value, we describe here experimental conditions allowing the production of large quantities (up to 35 mg/l of bacterial culture) of correctly refolded and totally functional single chain fragment variable (scFv). These quantities were sufficient to characterize the binding properties of this small recombinant fragment through in vitro and ex vivo approaches. Interestingly, this scFv retains full binding capacity for its antigen, i.e. the human prion protein, when compared with the corresponding Fab or whole antibody, and recognizes soluble, solid-phase-adsorbed, and membrane-bound prion protein. This strongly suggests that from the mAb cloning step to the refolding of the recombinant fragment, each stage is well controlled, leading to almost 100% functional scFv. These results are of interest not only in view of possible immunotherapy for prion diseases, but also more generally in emphasizing the great promise of these small recombinant molecules in the context of targeted therapies.
Databáze: OpenAIRE
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