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Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
Published on: August 27, 2021
Rapid prediction of expression and refolding yields using phage display
Kip Dudgeon1, Romain Rouet, Daniel Christ
1Garvan Institute of Medical Research, 384 Victoria Street, Darlinghurst, Sydney, NSW 2010, Australia.
Protein Engineering, Design & Selection : PEDS
|May 22, 2013
Summary
Researchers developed a robust phage-based heat-denaturation method to predict protein aggregation. This method accurately forecasts expression and refolding yields for recombinant antibody variable domains, improving protein production.
Area of Science:
- Biotechnology
- Protein Engineering
- Immunology
Background:
- Protein aggregation is a major challenge in recombinant protein production, limiting the yield of valuable therapeutic proteins.
- Controlling protein aggregation is crucial for efficient manufacturing of recombinant antibodies.
Purpose of the Study:
- To validate a previously reported heat-denaturation method on phage for assessing protein aggregation.
- To evaluate the method's robustness and its ability to predict protein behavior in solution.
Main Methods:
- Utilized a panel of human antibody variable heavy domains with defined aggregation propensities.
- Applied a heat-denaturation method on phage to assess protein stability and aggregation tendency.
Main Results:
- The heat-denaturation method on phage demonstrated robustness across various denaturation conditions.
- The method proved highly indicative of protein solution behavior.
- It served as an excellent predictor of both expression and refolding yields for antibody domains.
Conclusions:
- The phage-based heat-denaturation assay is a reliable tool for predicting protein aggregation and optimizing recombinant protein production.
- This validated method can guide protein engineering efforts to enhance the manufacturability of therapeutic antibodies.
