Selection and characterization of single chain Fv fragments against murine recombinant prion protein from a synthetic human antibody phage display library.
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 microM.
Leclerc
E
E
Institute of Molecular Biology and Biophysics, ETH Honggerberg, Zürich, Switzerland. estelle@scripps.edu
Liemann
S
S
Wildegger
G
G
Vetter
S W
SW
Nilsson
F
F
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D022801:Complementarity Determining Regions / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D015723:Gene Library; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology* / Q000302:isolation & purification*; D051379:Mice; D011328:Prions / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*
D022801:Complementarity Determining Regions; D007128:Immunoglobulin Fragments; D011328:Prions; D011994:Recombinant Proteins; C055966:immunoglobulin Fv