title: Target-specific NMR detection of protein-ligand interactions with antibody-relayed15N-group selective STD.
abstract: Fragment-based drug design has been successfully applied to challenging targets where the detection of the weak protein-ligand interactions is a key element.1H saturation transfer difference (STD) NMR spectroscopy is a powerful technique for this work but it requires pure homogeneous proteins as targets. Monoclonal antibody (mAb)-relayed15N-GS STD spectroscopy has been developed to resolve the problem of protein mixtures and impure proteins. A15N-labelled target-specific mAb is selectively irradiated and the saturation is relayed through the target to the ligand. Tests on the anti-Gal-1 mAb/Gal-1/lactose system showed that the approach is experimentally feasible in a reasonable time frame. This method allows detection and identification of binding molecules directly from a protein mixture in a multicomponent system.
authors.affiliation: Department of Medical Chemistry, University of Szeged, Dóm t. 8., Szeged, 6720, Hungary.
authors.forename: Anasztázia
authors.initials: A
authors.lastname: Hetényi
authors.affiliation: SZTE-MTA Lendület Foldamer Research Group, Institute of Pharmaceutical Analysis Department, University of Szeged, Somogyi u. 4, Szeged, 6720, Hungary.
authors.forename: Zsófia
authors.initials: Z
authors.lastname: Hegedűs
authors.affiliation: Lymphocyte Signal Transduction Laboratory, Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Temesvári krt. 62, Szeged, 6726, Hungary.
authors.forename: Roberta
authors.initials: R
authors.lastname: Fajka-Boja
authors.affiliation: Lymphocyte Signal Transduction Laboratory, Institute of Genetics, Biological Research Center of the Hungarian Academy of Sciences, Temesvári krt. 62, Szeged, 6726, Hungary.
authors.forename: Éva
authors.initials: É
authors.lastname: Monostori
authors.affiliation: Department of Inorganic and Analytical Chemistry, University of Debrecen, Egyetem tér 1., Debrecen, 4032, Hungary.
authors.forename: Katalin E
authors.initials: KE
authors.lastname: Kövér
authors.affiliation: SZTE-MTA Lendület Foldamer Research Group, Institute of Pharmaceutical Analysis Department, University of Szeged, Somogyi u. 4, Szeged, 6720, Hungary. martinek@pharm.u-szeged.hu.
authors.forename: Tamás A
authors.initials: TA
authors.lastname: Martinek
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D015195:Drug Design; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008958:Models, Molecular; D008968:Molecular Conformation; D009587:Nitrogen Isotopes / Q000737:chemistry*; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D008024:Ligands; D009587:Nitrogen Isotopes; C000615243:Nitrogen-15; D011506:Proteins
keywords: Antibody; GS-STD; Gal-1; Protein mixture; Protein–ligand interaction