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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 HubbuchEmail author
Research Paper
  • 74 Downloads

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

Keywords

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

Notes

Acknowledgements

This work was financially supported by the BE-Basic Foundation (https://www.be-basic.org), under project FS2.003. The authors want to thank academic and industrial partners for scientific discussions during the development of this work. Additionally, the authors would like to thank Lena Enghauser for contributing to the experimental work.

Compliance with ethical standards

Conflict of interest

The authors report no conflict of interest.

Supplementary material

449_2019_2239_MOESM1_ESM.pdf (1.2 mb)
Supplementary material 1 (PDF 1262 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Marieke E. Klijn
    • 1
  • Anna K. Wöll
    • 1
  • Jürgen Hubbuch
    • 1
    Email author
  1. 1.Institute of Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT)KarlsruheGermany

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