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Monoclonal antibody stability can be usefully monitored using the excitation-energy-dependent fluorescence edge-shift.
Knight, Michael J; Woolley, Rachel E; Kwok, Anthony; Parsons, Stuart; Jones, Hannah B L; Gulácsy, Christina E; Phaal, Polly; Kassaar, Omar; Dawkins, Kieran; Rodriguez, Elizabeth; Marques, Andreia; Bowsher, Leo; Wells, Stephen A; Watts, Andrew; van den Elsen, Jean M H; Turner, Alison; O'Hara, John; Pudney, Christopher R.
Afiliación
  • Knight MJ; UCB, 216 Bath Road, Slough SL1 3WE, U.K.
  • Woolley RE; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Kwok A; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Parsons S; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Jones HBL; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Gulácsy CE; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Phaal P; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Kassaar O; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • Dawkins K; UCB, 216 Bath Road, Slough SL1 3WE, U.K.
  • Rodriguez E; UCB, 216 Bath Road, Slough SL1 3WE, U.K.
  • Marques A; UCB, 216 Bath Road, Slough SL1 3WE, U.K.
  • Bowsher L; UCB, 216 Bath Road, Slough SL1 3WE, U.K.
  • Wells SA; Department of Physics, University of Bath, Bath, U.K.
  • Watts A; Department of Pharmacy and Pharmacology, University of Bath, Bath, U.K.
  • van den Elsen JMH; Centre for Therapeutic Innovation, University of Bath, Bath, U.K.
  • Turner A; Department of Biology and Biochemistry, University of Bath, Bath, U.K.
  • O'Hara J; Centre for Therapeutic Innovation, University of Bath, Bath, U.K.
  • Pudney CR; UCB, 216 Bath Road, Slough SL1 3WE, U.K.
Biochem J ; 477(18): 3599-3612, 2020 09 30.
Article en En | MEDLINE | ID: mdl-32869839
Among the major challenges in the development of biopharmaceuticals are structural heterogeneity and aggregation. The development of a successful therapeutic monoclonal antibody (mAb) requires both a highly active and also stable molecule. Whilst a range of experimental (biophysical) approaches exist to track changes in stability of proteins, routine prediction of stability remains challenging. The fluorescence red edge excitation shift (REES) phenomenon is sensitive to a range of changes in protein structure. Based on recent work, we have found that quantifying the REES effect is extremely sensitive to changes in protein conformational state and dynamics. Given the extreme sensitivity, potentially this tool could provide a 'fingerprint' of the structure and stability of a protein. Such a tool would be useful in the discovery and development of biopharamceuticals and so we have explored our hypothesis with a panel of therapeutic mAbs. We demonstrate that the quantified REES data show remarkable sensitivity, being able to discern between structurally identical antibodies and showing sensitivity to unfolding and aggregation. The approach works across a broad concentration range (µg-mg/ml) and is highly consistent. We show that the approach can be applied alongside traditional characterisation testing within the context of a forced degradation study (FDS). Most importantly, we demonstrate the approach is able to predict the stability of mAbs both in the short (hours), medium (days) and long-term (months). The quantified REES data will find immediate use in the biopharmaceutical industry in quality assurance, formulation and development. The approach benefits from low technical complexity, is rapid and uses instrumentation which exists in most biochemistry laboratories without modification.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Anticuerpos Monoclonales Idioma: En Revista: Biochem J Año: 2020 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Anticuerpos Monoclonales Idioma: En Revista: Biochem J Año: 2020 Tipo del documento: Article Pais de publicación: Reino Unido