TY  - JOUR
AU  - Müller-Schiffmann, A.
AU  - Petsch, B.
AU  - Leliveld, S. R.
AU  - Muyrers, J.
AU  - Salwierz, A.
AU  - Mangels, Ch.
AU  - Schwarzinger, St.
AU  - Riesner, D.
AU  - Stitz, L.
AU  - Korth, C.
TI  - Complementarity determining regions of an anti-prion protein scFv fragment orchestrate conformation specificity and antiprion activity
JO  - Molecular immunology
VL  - 46
SN  - 0161-5890
CY  - Oxford
PB  - Pergamon Press
M1  - PreJuSER-2682
SP  - 532 - 540
PY  - 2009
N1  - This work was funded by a grant from the Forschungskommission of the University of Dusseldorf Medical School (to R.L.), a grant from the VW foundation (to CX and S.S.), a grant from the TSE-Program Baden-Wuerttemberg (729.59-7/1; to L.S.), a grant from the DFG (GRK1033; to CK), and a grant from the FP6, EU Research Commission Anteprion (LSHB-C7-2006-019090; to CX and L.S.).
AB  - The prion protein, PrP, exists in several stable conformations, with the presence of one conformation, PrP(Sc), associated with transmissible neurodegenerative diseases. Targeting PrP by high-affinity ligands has been proven to be an effective way of preventing peripheral prion infections. Here, we have generated bacterially expressed single chain fragments of the variable domains (scFv) of a monoclonal antibody in Escherichia coli, originally raised against purified PrP(Sc) that recognizes both PrP(C) and PrP(Sc). This scFv fragment had a dissociation constant (K(D)) with recombinant PrP of 2 nM and cleared prions in ScN2a cells at 4 nM, as demonstrated by a mouse prion bioassay. A peptide corresponding to the complementarity determining region 3 of the heavy chain (CDR3H) selectively bound PrP(Sc) but had lost antiprion activity. However, synthesis and application of an improved peptide mimicking side chain topology of CDR3H while exhibiting increased protease resistance, a retro-inverso d-peptide of CDR3H, still bound PrP(Sc) and reinstated antiprion activity. We conclude that (1) scFvW226 is so far the smallest polypeptide with bioassay confirmed antiprion activity, and (2) differential conformation specificity and bioactivity can be regulated by orchestrating the participation of different CDRs.
KW  - Amino Acid Sequence
KW  - Animals
KW  - Antibody Affinity: immunology
KW  - Cell Line
KW  - Complementarity Determining Regions: immunology
KW  - Complementarity Determining Regions: metabolism
KW  - Mice
KW  - Mice, Knockout
KW  - Molecular Sequence Data
KW  - Peptides: chemistry
KW  - Peptides: immunology
KW  - Peptides: metabolism
KW  - PrPC Proteins: immunology
KW  - PrPC Proteins: metabolism
KW  - PrPSc Proteins: immunology
KW  - PrPSc Proteins: metabolism
KW  - Protein Conformation
KW  - Complementarity Determining Regions (NLM Chemicals)
KW  - Peptides (NLM Chemicals)
KW  - PrPC Proteins (NLM Chemicals)
KW  - PrPSc Proteins (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:18973947
UR  - <Go to ISI:>//WOS:000263429500004
DO  - DOI:10.1016/j.molimm.2008.07.023
UR  - https://juser.fz-juelich.de/record/2682
ER  -