Isothermal chemical denaturation as a complementary tool to overcome limitations of thermal differential scanning fluorimetry in predicting physical stability of protein formulations

Svilenov, Hristo ; Markoja, Uros und Winter, Gerhard (April 2018): Isothermal chemical denaturation as a complementary tool to overcome limitations of thermal differential scanning fluorimetry in predicting physical stability of protein formulations. In: European Journal of Pharmaceutics and Biopharmaceutics, Bd. 125: S. 106-113

Abstract

Various stability indicating techniques find application in the early stage development of novel therapeutic protein candidates. Some of these techniques are used to select formulation conditions that provide high protein physical stability. Such approach is highly dependent on the reliability of the stability indicating technique used. In this work, we present a formulation case study in which we evaluate the ability of differential scanning fluorimetry (DSF) and isothermal chemical denaturation (ICD) to predict the physical stability of a model monoclonal antibody during accelerated stability studies. First, we show that a thermal denaturation technique like DSF can provide misleading physical stability rankings due to buffer specific pH shifts during heating. Next, we demonstrate how isothermal chemical denaturation can be used to tackle the above-mentioned challenge. Subsequently, we show that the concentration dependence of the Gibbs free energy of unfolding determined by ICD provides better predictions for the protein physical stability in comparison to the often-used Tm (melting temperature of the protein determined with DSF) and Cm (concentration of denaturant needed to unfold 50% of the protein determined with ICD). Finally, we give a suggestion for a rational approach which includes a combination of DSF and ICD to obtain accurate and reliable protein physical stability ranking in different formulations.

• Dokumententyp: Zeitschriftenartikel
• EU Funded Grant Agreement Number: 675074
• EU-Projekte: Horizon 2020 > Marie Skłodowska Curie Actions > Marie Skłodowska-Curie Innovative Training Networks > 675074: PIPPI - Protein-excipient Interactions and Protein-Protein Interactions in formulation
• Publikationsform: Postprint
• Keywords: Protein formulation; Thermal denaturation; Isothermal chemical denaturation; Monoclonal antibody; Differential scanning fluorimetry
• Fakultät: Chemie und Pharmazie > Department für Pharmazie - Zentrum für Pharmaforschung
• Themengebiete: 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie
• ISSN: 0939-6411
• Sprache: Englisch

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