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A generic hierarchical screening method for the analysis of microscale refolds using an automated robotic platform
Gemma C Ordidge1, Gareth Mannall, John Liddell
1Dept. of Biochemical Engineering, Advanced Centre for Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE. g.ordidge@ucl.ac.uk
Biotechnology Progress
|January 17, 2012
Summary
Optimizing protein refolding from inclusion bodies is accelerated using an automated platform with hierarchical assays. This approach efficiently screens conditions, improving yields and reducing process development time.
Area of Science:
- Biotechnology
- Protein Chemistry
- Process Development
Background:
- Protein refolding from inclusion bodies often results in low active protein yields, hindering biopharmaceutical development.
- Empirical optimization of refolding conditions is time-consuming and resource-intensive, slowing down process development timelines.
Purpose of the Study:
- To develop an automated, microscale platform for high-throughput screening of protein refolding conditions.
- To implement a hierarchical assay strategy to efficiently identify optimal refolding parameters, saving resources and maximizing information output.
Main Methods:
- An automated robotic platform was utilized to create a dilution refolding process-screening system.
- A hierarchical assay approach was employed, starting with absorbance and fluorescence assays, followed by activity or RP-HPLC assays.
- The platform was used to study and minimize the impact of oxygenation on redox potential and misfolding.
Main Results:
- Hierarchical assays, combining absorbance and fluorescence, effectively screened out low-yielding conditions, significantly reducing the number of samples for further analysis.
- This method identified optimal refolding conditions more rapidly and efficiently compared to traditional empirical approaches.
- The platform demonstrated its capability to accelerate data acquisition for process development in a resource-efficient manner.
Conclusions:
- Automated, hierarchical assay screening is a powerful strategy to optimize protein refolding from inclusion bodies.
- This approach significantly reduces bottlenecks in high-throughput analysis, speeding up process development.
- The developed platform enables efficient data collection for evaluating diverse refolding conditions, enhancing resource utilization.
