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Apparent protein cloud point temperature determination using a low volume high-throughput cryogenic device in
Marieke E Klijn1, Anna K Wöll1, Jürgen Hubbuch2
1Institute of Engineering in Life Sciences, Section IV: Biomolecular Separation Engineering, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 2, 76131, Karlsruhe, Germany.
Bioprocess and Biosystems Engineering
|November 23, 2019
Summary
This study introduces a novel, low-volume, high-throughput method for determining the cloud point temperature of proteins. This technique aids in predicting long-term protein stability, enhancing formulation development efficiency.
Area of Science:
- Biochemistry
- Protein Science
- Formulation Development
Background:
- Protein stability is crucial for product development.
- Predicting long-term protein stability requires efficient short-term parameters.
- The protein cloud point temperature (Tcloud) is a key parameter.
Purpose of the Study:
- To develop a low-volume, high-throughput method for sub-zero Tcloud determination.
- To establish a method for lysozyme Tcloud measurement.
- To correlate Tcloud with long-term protein stability.
Main Methods:
- Utilized a cryogenic device with an automated imaging system.
- Developed a low-volume, high-throughput detection method.
- Performed robustness and validation studies.
Main Results:
- Achieved high reproducibility (0.2 °C median absolute deviation).
- Validated results against literature (Pearson correlation coefficient of 0.996).
- Demonstrated partial correlation between Tcloud and long-term lysozyme stability under various conditions.
Conclusions:
- The developed method enables efficient, low-volume Tcloud determination.
- This approach can advance short-term strategies for protein formulation.
- The Tcloud parameter shows potential for predicting protein stability.
