Chemometrics in Protein Formulation: Stability Governed by Repulsion and Protein Unfolding

Research output: Contribution to journalJournal articleResearchpeer-review

  • Alina Kulakova
  • Dillen Augustijn
  • Inas El Bialy
  • Lorenzo Gentiluomo
  • Maria Laura Greco
  • Stefan Hervø-hansen
  • Sowmya Indrakumar
  • Sujata Mahapatra
  • Marcello Martinez Morales
  • Christin Pohl
  • Marco Polimeni
  • Aisling Roche
  • Hristo L. Svilenov
  • Andreas Tosstorff
  • Matja Zalar
  • Robin Curtis
  • Jeremy P. Derrick
  • Wolfgang Frieß
  • Alexander P. Golovanov
  • Mikael Lund
  • Allan Nørgaard
  • Tarik A. Khan
  • Günther H. J. Peters
  • Alain Pluen
  • Dierk Roessner
  • Werner W. Streicher
  • Christopher F. Van Der Walle
  • Jim Warwicker
  • Shahid Uddin
  • Gerhard Winter
  • Jens Thostrup Bukrinski
  • Pernille Harris
Therapeutic proteins can be challenging to develop due to their complexity and the requirement of an acceptable formulation to ensure patient safety and efficacy. To date, there is no universal formulation development strategy that can identify optimal formulation conditions for all types of proteins in a fast and reliable manner. In this work, high-throughput characterization, employing a toolbox of five techniques, was performed on 14 structurally different proteins formulated in 6 different buffer conditions and in the presence of 4 different excipients. Multivariate data analysis and chemometrics were used to analyze the data in an unbiased way. First, observed changes in stability were primarily determined by the individual protein. Second, pH and ionic strength are the two most important factors determining the physical stability of proteins, where there exists a significant statistical interaction between protein and pH/ionic strength. Additionally, we developed prediction methods by partial least-squares regression. Colloidal stability indicators are important for prediction of real-time stability, while conformational stability indicators are important for prediction of stability under accelerated stress conditions at 40 °C. In order to predict real-time storage stability, protein–protein repulsion and the initial monomer fraction are the most important properties to monitor.
Original languageEnglish
JournalMolecular Pharmaceutics
Issue number6
Pages (from-to)2951-2965
Number of pages15
Publication statusPublished - 2023

ID: 357475998