Journal Article

PreJuSER-2682

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Complementarity determining regions of an anti-prion protein scFv fragment orchestrate conformation specificity and antiprion activity

Müller-Schiffmann, A. ; Petsch, B. ; Leliveld, S. R.FZJ* ; Muyrers, J. ; Salwierz, A. ; Mangels, C. ; Schwarzinger, S. ; Riesner, D. ; Stitz, L. ; Korth, C.

2009
Pergamon Press Oxford

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Please use a persistent id in citations: doi:10.1016/j.molimm.2008.07.023

Abstract: 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.

Keyword(s): Amino Acid Sequence (MeSH) ; Animals (MeSH) ; Antibody Affinity: immunology (MeSH) ; Cell Line (MeSH) ; Complementarity Determining Regions: immunology (MeSH) ; Complementarity Determining Regions: metabolism (MeSH) ; Mice (MeSH) ; Mice, Knockout (MeSH) ; Molecular Sequence Data (MeSH) ; Peptides: chemistry (MeSH) ; Peptides: immunology (MeSH) ; Peptides: metabolism (MeSH) ; PrPC Proteins: immunology (MeSH) ; PrPC Proteins: metabolism (MeSH) ; PrPSc Proteins: immunology (MeSH) ; PrPSc Proteins: metabolism (MeSH) ; Protein Conformation (MeSH) ; Complementarity Determining Regions ; Peptides ; PrPC Proteins ; PrPSc Proteins ; J ; Prions (auto) ; Monoclonal antibody (auto) ; Conformation specificity (auto) ; Complementarity determining regions (auto) ; Recombinant antibodies (auto) ; Retro-inverso D-peptides (auto)

Note: 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.).

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Contributing Institute(s):

1. Molekulare Biophysik (INB-2)

Research Program(s):

1. Funktion und Dysfunktion des Nervensystems (P33)

Appears in the scientific report 2010

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