title: By-passing large screening experiments using sequencing as a tool to identify scFv fragments targeting atherosclerotic lesions in a novel in vivo phage display selection.
abstract: Atherosclerosis is a chronic, progressive inflammatory disease that may develop into vulnerable lesions leading to thrombosis. To interrogate the molecular components involved in this process, single-chain variable fragments (scFvs) from a semi-synthetic human antibody library were selected on the lesions induced in a rabbit model of atherosclerosis after two rounds of in vivo phage display. Homing Phage-scFvs were isolated from (1) the injured endothelium, (2) the underlying lesional tissue and (3) the cells within the intima. Clones selected on the basis of their redundancy or the presence of key amino acids, as determined by comparing the distribution between the native and the selected libraries, were produced in soluble form, and seven scFvs were shown to specifically target the endothelial cell surface and inflamed intima-related regions of rabbit tissue sections by immunohistology approaches. The staining patterns differed depending on the scFv compartment of origin. This study demonstrates that large-scale scFv binding assays can be replaced by a sequence-based selection of best clones, paving the way for easier use of antibody libraries in in vivo biopanning experiments. Future investigations will be aimed at characterizing the scFv/target couples by mass spectrometry to set the stage for more accurate diagnostic of atherosclerosis and development of therapeutic strategies.
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.|Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France.
authors: Kamel
authors: K
authors: Deramchia
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.|Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France.
authors: Marie-Josee
authors: MJ
authors: Jacobin-Valat
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.|Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France.
authors: Jeanny
authors: J
authors: Laroche-Traineau
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.|Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France.
authors: Stephane
authors: S
authors: Bonetto
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.
authors: Stephane
authors: S
authors: Sanchez
authors: Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France.
authors: Pierre
authors: P
authors: Dos Santos
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.
authors: Philippe
authors: P
authors: Massot
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.
authors: Jean-Michel
authors: JM
authors: Franconi
authors: MCRI, Montpellier Cancer Research Institute , INSERM, U896, Montpellier1 University, CRLC Val d'Aurelle Paul Lamarque, Montpellier, F-34298, France.
authors: Pierre
authors: P
authors: Martineau
authors: Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France.|Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France.
authors: Gisele
authors: G
authors: Clofent-Sanchez
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001012:Aorta, Abdominal / Q000473:pathology; D050197:Atherosclerosis / Q000503:physiopathology* / Q000628:therapy*; D005487:Foam Cells / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation / Q000473:pathology; D008279:Magnetic Resonance Imaging; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D057127:Single-Chain Antibodies / Q000737:chemistry*; D017539:Tunica Intima / Q000473:pathology
chemical_list: D019151:Peptide Library; D057127:Single-Chain Antibodies