NR AVVL
AU Adamson,C.S.; Yao,Y.; Vasiljevic,S.; Sy,M.S.; Ren,J.; Jones,I.M.
TI Novel single chain antibodies to the prion protein identified by phage display
QU Virology 2007 Feb 5; 358(1): 166-77
PT journal article; research support, non-u.s. gov't
AB A well defined structure is available for the carboxyl half of the cellular prion protein (PrPc), while the structure of the amino terminal half of the molecule remains ill defined. The unstructured nature of the polypeptide has meant that relatively few of the many antibodies generated against PrPc recognise this region. To circumvent this problem, we have used a previously characterised and well expressed fragment derived from the amino terminus of PrPc as bait for panning a single chain antibody phage (scFv-P) library. Using this approach, we identified and characterised 1 predominant and 3 additional scFv-Ps that contained different V(H) and V(L) sequences and that bound specifically to the PrPc target. Epitope mapping revealed that all scFv-Ps recognised linear epitopes between PrPc residues 76 and 156. When compared with existing monoclonal antibodies (MAb), the binding of the scFvs was significantly different in that high level binding was evident on truncated forms of PrPc that reacted poorly or not at all with several pre-existing MAbs. These data suggest that the isolated scFv-Ps bind to novel epitopes within the amino-central region of PrPc. In addition, the binding of MAbs to known linear epitopes within PrPc depends strongly on the endpoints of the target PrPc fragment used.
MH Amino Acid Sequence; Antibodies/*immunology/*isolation & purification; Antibodies, Monoclonal/immunology; Antibody Affinity; Antibody Specificity; Binding Sites, Antibody; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Immunoglobulin Fragments/*immunology/*isolation & purification; Immunoglobulin Variable Region; *Peptide Library; PrPc Proteins/chemistry/*immunology; Protein Structure, Tertiary; Sequence Alignment
AD School of Biological Sciences, The University of Reading, Reading, RG6 6AJ, UK
SP englisch
PO USA