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Determination of Binding Sites on Trastuzumab and Pertuzumab to Selective Affimers Using Hydrogen-Deuterium Exchange Mass Spectrometry.
Olaleye, Oladapo; Graf, Christian; Spanov, Baubek; Govorukhina, Natalia; Groves, Matthew R; van de Merbel, Nico C; Bischoff, Rainer.
Afiliación
  • Olaleye O; Analytical Biochemistry, Department of Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
  • Graf C; Novartis Technical Research & Development Biologics, Hexal AG, Keltenring 1 + 3, 82041 Oberhaching, Germany.
  • Spanov B; Analytical Biochemistry, Department of Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
  • Govorukhina N; Analytical Biochemistry, Department of Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
  • Groves MR; Drug Design, Department of Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
  • van de Merbel NC; Analytical Biochemistry, Department of Pharmacy, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
  • Bischoff R; ICON Bioanalytical Laboratories, Amerikaweg 18, 9407 TK Assen, The Netherlands.
J Am Soc Mass Spectrom ; 34(4): 775-783, 2023 Apr 05.
Article en En | MEDLINE | ID: mdl-36960982
Hydrogen-deuterium exchange mass spectrometry (HDX-MS) is a method to probe the solvent accessibility and conformational dynamics of a protein or a protein-ligand complex with respect to exchangeable amide hydrogens. Here, we present the application of HDX-MS to determine the binding sites of Affimer reagents to the monoclonal antibodies trastuzumab and pertuzumab, respectively. Intact and subunit level HDX-MS analysis of antibody-affimer complexes showed significant protection from HDX in the antibody Fab region upon affimer binding. Bottom-up HDX-MS experiments including online pepsin digestion revealed that the binding sites of the affimer reagents were mainly located in the complementarity-determining region (CDR) 2 of the heavy chain of the respective antibodies. Three-dimensional models of the binding interaction between the affimer reagents and the antibodies were built by homology modeling and molecular docking based on the HDX data.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Medición de Intercambio de Deuterio / Espectrometría de Masas de Intercambio de Hidrógeno-Deuterio Idioma: En Revista: J Am Soc Mass Spectrom Año: 2023 Tipo del documento: Article País de afiliación: Países Bajos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Medición de Intercambio de Deuterio / Espectrometría de Masas de Intercambio de Hidrógeno-Deuterio Idioma: En Revista: J Am Soc Mass Spectrom Año: 2023 Tipo del documento: Article País de afiliación: Países Bajos
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