Apparent protein cloud point temperature determination using a low volume high-throughput cryogenic device in combination with automated imaging

Marieke E. Klijn, Anna K. Wöll, Jürgen Hubbuch

Research output: Contribution to journalArticleScientificpeer-review

3 Citations (Scopus)

Abstract

Abstract: Short-term parameters correlating to long-term protein stability, such as the protein cloud point temperature (Tcloud), are of interest to improve efficiency during protein product development. Such efficiency is reached if short-term parameters are obtained in a low volume and high-throughput (HT) manner. This study presents a low volume HT detection method for (sub-zero) Tcloud determination of lysozyme, as such an experimental method is not available yet. The setup consists of a cryogenic device with an automated imaging system. Measurement reproducibility (median absolute deviation of 0.2 °C) and literature-based parameter validation (Pearson correlation coefficient of 0.996) were shown by a robustness and validation study. The subsequent case study demonstrated a partial correlation between the obtained apparent Tcloud parameter and long-term protein stability as a function of lysozyme concentration, ion type, ionic strength, and freeze/thaw stress. The presented experimental setup demonstrates its ability to advance short-term strategies for efficient protein formulation development. Graphical Abstract: [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)439-456
JournalBioprocess and Biosystems Engineering
Volume43
Issue number3
DOIs
Publication statusPublished - 2019
Externally publishedYes

Keywords

  • Freeze/thaw
  • Liquid–liquid phase separation
  • Long-term protein stability
  • Lysozyme
  • Phase diagrams

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