Int. J. Mol. Sci. 2012, 13(6), 6902-6923; doi:10.3390/ijms13066902
Article

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

1 Magnetic Resonance Center of Biological Systems, UMR 5536, National Center for Scientific Research, Bordeaux Segalen University, 33076 Bordeaux Cedex, France 2 Technology Platform for Biomedical Innovation, Bordeaux Segalen University, 33600 Bordeaux Cedex, France 3 MCRI, Montpellier Cancer Research Institute , INSERM, U896, Montpellier1 University, CRLC Val d’Aurelle Paul Lamarque, Montpellier, F-34298, France These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 27 March 2012; in revised form: 11 May 2012 / Accepted: 22 May 2012 / Published: 7 June 2012
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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.
Keywords: atherosclerosis; human antibody; single-chain variable fragment (scFv); semi-synthetic phage display library; in vivo phage display; biopanning

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MDPI and ACS Style

Deramchia, K.; Jacobin-Valat, M.-J.; Laroche-Traineau, J.; Bonetto, S.; Sanchez, S.; Dos Santos, P.; Massot, P.; Franconi, J.-M.; Martineau, P.; Clofent-Sanchez, G. 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. Int. J. Mol. Sci. 2012, 13, 6902-6923.

AMA Style

Deramchia K, Jacobin-Valat M-J, Laroche-Traineau J, Bonetto S, Sanchez S, Dos Santos P, Massot P, Franconi J-M, Martineau P, Clofent-Sanchez G. 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. International Journal of Molecular Sciences. 2012; 13(6):6902-6923.

Chicago/Turabian Style

Deramchia, Kamel; Jacobin-Valat, Marie-Josee; Laroche-Traineau, Jeanny; Bonetto, Stephane; Sanchez, Stephane; Dos Santos, Pierre; Massot, Philippe; Franconi, Jean-Michel; Martineau, Pierre; Clofent-Sanchez, Gisele. 2012. "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." Int. J. Mol. Sci. 13, no. 6: 6902-6923.

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