(1170-C) Complete Workflow Automation for High-Throughput Western Blotting in Drug Discovery

Abstract: The ProteinSimpleTM Jess system for automated western blotting is utilized in drug discovery applications like targeted protein degradation (TPD). In this study, protein quantification and Jess plate preparation were performed with the CyBio FeliX pipetting robot from Analytik Jena. PROTAC THAL-SNS-032 and its target molecule CDK-9 were used to demonstrate the automation capabilities. First, the CyBio FeliX performed serial dilutions of the cell lysates and filled the Jess plate with different antibody dilutions for the optimization of the anti-CDK9 antibody. Then, the liquid handler was used to prepare a BCA assay to determine the protein concentration in lysates of PROTAC-treated cells. Last, antibody optimization results and BCA assay data were combined to set up the Jess plates, again using the CyBio FeliX liquid handler for process automation.
Optimization of the anti-CDK9 antibody ensured saturation and linearity of the signal and enabled quantifiable data. Data analysis confirmed linear, protein concentration-dependent detection of CDK9 and consistent signals indicating saturation across all antibody dilutions. The dose-dependent CDK9 degradation triggered by SNS-THAL-032 remained constant across multiple experiments.
The automated process executed by the CyBio FeliX liquid handler demonstrated equivalent quality to the manual process, offering a time-saving advantage of up to 60 minutes per Jess run. In summary, the study illustrates the utility of the CyBio FeliX in optimizing antibody and sample protein concentrations, streamlining the preparation of ProteinSimpleTM Jess plates for efficient screening purposes.