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Harnessing Liquid Crystal Sensors for High-Throughput Real-Time Detection of Structural Changes in Lysozyme during
Lili Guo1, Jing Zhao1, Zongfu An1
1School of Chemical Engineering, Sungkyunkwan University (SKKU), Suwon 16419, Republic of Korea.
Analytical Chemistry
|November 16, 2023
Summary
A novel liquid crystal sensor offers real-time monitoring of protein refolding efficiency. This technology overcomes limitations of traditional methods, enabling faster optimization of refolding conditions for recombinant proteins.
Area of Science:
- Biophysical Chemistry
- Materials Science
- Biotechnology
Background:
- Traditional protein refolding assessment relies on slow, off-line assays like HPLC.
- This time lag hinders optimization of refolding conditions, especially for recombinant proteins expressed as inclusion bodies.
- A need exists for rapid, real-time methods to monitor protein conformational changes during refolding.
Purpose of the Study:
- To develop and demonstrate a novel liquid crystal (LC)-based sensor for real-time assessment of protein refolding efficiency.
- To utilize the sensor's ability to detect structural changes during protein unfolding and refolding.
- To provide a versatile platform for optimizing protein refolding processes.
Main Methods:
- Fabrication of a sensing layer using 5CB liquid crystals intercalated with DOPE phospholipid.
- Utilizing changes in DOPE-lipid alignment at the aqueous/LC interface due to protein surface charge fluctuations.
- Monitoring alterations in LC molecular ordering (homeotropic/planar transitions) via polarized optical microscopy and grayscale analysis.
- Demonstrating proof-of-concept using lysozyme as a model protein.
Main Results:
- The LC sensor successfully detected conformational changes in lysozyme during unfolding and refolding.
- Changes in LC layer brightness, quantified by grayscale analysis, correlated directly with lysozyme structural transitions.
- The sensor demonstrated compatibility with various refolding buffer conditions, enabling real-time monitoring.
- The system effectively discriminated between different protein structural states.
Conclusions:
- A novel LC-based sensor provides a sensitive and real-time method for evaluating protein refolding efficiency.
- This technology overcomes the limitations of traditional off-line assays, facilitating faster optimization of protein refolding.
- The sensor holds significant potential for studies involving diverse proteins, particularly those lacking established activity assays.
