Selection and characterization of single chain Fv fragments against murine recombinant prion protein from a synthetic human antibody phage display library

Article type: Research Article

Authors: Leclerc, Estelle^{a; *} | Liemann, Susanne^a | Wildegger, Gudrun^a | Vetter, Stefan W.^b | Nilsson, Fredrik^a

Affiliations: [a] Institute of Molecular Biology and Biophysics, ETH Honggerberg, Zürich, Switzerland | [b] Institute of Biochemistry III, ETH-Zentrum, Zürich, Switzerland

Correspondence: [*] Corresponding author: Estelle Leclerc, Department of Immunology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA. Tel.: +1 858 784 9663; Fax: +1 858 784 8360; E-mail: [email protected].

Abstract: We describe the selection of single chain Fv fragments (scFv) against recombinant murine prion protein (mPrP) from a synthetic human antibody phage display library. Six different antibodies were isolated after three rounds of panning against full-length mPrP. All antibodies recognized a truncated form of mPrP containing residues (121–231). The amino acid sequence of the CDR3 of the scFv fragments has been determined. Five of the antibodies have been over-expressed, purified and their affinity for full-length mPrP determined by ELISA. The observed binding affinities vary from 30 nM to 2.7 µM.

DOI: 10.3233/HAB-2000-9403

Journal: Human Antibodies, vol. 9, no. 4, pp. 207-214, 2000