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Understanding the pathway and kinetics of aspartic acid isomerization in peptide mapping methods for monoclonal antibodies.
Kuang, June; Tao, Yuanqi; Song, Yuanli; Chemmalil, Letha; Mussa, Nesredin; Ding, Julia; Li, Zheng Jian.
Afiliación
  • Kuang J; Biologics Development Organization, Bristol-Myers Squibb Company, Devens, MA, 01434, USA.
  • Tao Y; Biologics Development Organization, Bristol-Myers Squibb Company, Devens, MA, 01434, USA.
  • Song Y; Analytical Science Biologics, Takeda Pharmaceutical Company, Lexington, MA, 02421, USA.
  • Chemmalil L; Biologics Development Organization, Bristol-Myers Squibb Company, Devens, MA, 01434, USA.
  • Mussa N; Process Development & Manufacture Operations, GSK, MA, 02451, Waltham, USA.
  • Ding J; Biologics Development Organization, Bristol-Myers Squibb Company, Devens, MA, 01434, USA.
  • Li ZJ; Biologics Development Organization, Bristol-Myers Squibb Company, Devens, MA, 01434, USA.
Anal Bioanal Chem ; 413(8): 2113-2123, 2021 Mar.
Article en En | MEDLINE | ID: mdl-33543314
Isomerization of aspartic acid (Asp) in therapeutic proteins could lead to safety and efficacy concerns. Thus, accurate quantitation of various Asp isomerization along with kinetic understanding of the variant formations is needed to ensure optimal process development and sufficient product quality control. In this study, we first observed Asp-succinimide conversion in complementarity-determining regions (CDRs) Asp-Gly motif of a recombinant mAb through ion exchange chromatography, intact protein analysis by mass spectrometry, and LC-MS/MS. Then, we developed a specific peptide mapping method, with optimized sample digestion conditions, to accurately quantitate Asp-succinimide-isoAsp variants at peptide level without method-induced isomerization. Various kinetics of Asp-succinimide-isoAsp isomerization pathways were elucidated using 18O labeling followed by LC-MS analysis. Molecular modeling and molecular dynamic simulation provide additional insight on the kinetics of Asp-succinimide formation and stability of succinimide intermediate. Findings of this work shed light on the molecular construct and the kinetics of the formation of isoAsp and succinimide in peptides and proteins, which facilitates analytical method development, protein engineering, and late phase development for commercialization of therapeutic proteins.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Péptidos / Mapeo Peptídico / Ácido Aspártico / Anticuerpos Monoclonales Tipo de estudio: Prognostic_studies Idioma: En Revista: Anal Bioanal Chem Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Alemania
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Péptidos / Mapeo Peptídico / Ácido Aspártico / Anticuerpos Monoclonales Tipo de estudio: Prognostic_studies Idioma: En Revista: Anal Bioanal Chem Año: 2021 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Alemania