title: The therapeutically anti-prion active antibody-fragment scFv-W226: paramagnetic relaxation-enhanced NMR spectroscopy aided structure elucidation of the paratope-epitope interface.
abstract: Antibodies have become indispensable reagents with numerous applications in biological and biotechnical analysis, in diagnostics as well as in therapy. In all cases, selective interaction with an epitope is crucial and depends on the conformation of the paratope. While epitopes are routinely mapped at high throughput, methods revealing structural insights on a rather short timescale are rare. We here demonstrate paramagnetic relaxation-enhanced (PRE) NMR spectroscopy to be a powerful tool unraveling structural information about epitope-orientation in a groove spanned by the complementary determining regions. In particular, we utilize the spin label TOAC, which is fused to the peptidic epitope using standard solid-phase chemistry and which is characterized by a reduced mobility compared to, e.g., spin labels attached to the side-chain functionalities of cysteine or lysine residues. We apply the method to determine the orientation of helix 1 of the prion protein, which is the epitope for the therapeutically anti-prion active scF(v) fragment W226.
authors.affiliation: Department of Biopolymers, Universitat Bayreuth, Universitatsstrasse 30, 95440 Bayreuth, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Mangels
authors.forename: Ruth
authors.initials: R
authors.lastname: Kellner
authors.forename: Jürgen
authors.initials: J
authors.lastname: Einsiedel
authors.forename: Philipp R
authors.initials: PR
authors.lastname: Weiglmeier
authors.forename: Paul
authors.initials: P
authors.lastname: Rosch
authors.forename: Peter
authors.initials: P
authors.lastname: Gmeiner
authors.forename: Stephan
authors.initials: S
authors.lastname: Schwarzinger
mesh_terms: D000595:Amino Acid Sequence; D001666:Binding Sites, Antibody; D000939:Epitopes; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011328:Prions / Q000737:chemistry* / Q000276:immunology; D011487:Protein Conformation; D016415:Sequence Alignment; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D013113:Spin Labels
chemical_list: D000939:Epitopes; D011328:Prions; D057127:Single-Chain Antibodies; D013113:Spin Labels