Detecting Secretory Proteins by Acoustic Droplet Ejection in Multiplexed High-Throughput Applications

Area of Science:

• Biochemistry
• Cell Biology
• Drug Discovery

Background:

• Secretory proteins are crucial for cell communication and implicated in various diseases.
• High-throughput detection methods are needed to identify therapies for secretion-related disorders.

Purpose of the Study:

• To develop a robust, multiplexed drug discovery platform for quantifying secreted proteins.
• To integrate extracellular protein analysis with intracellular and cytotoxicity assays.

Main Methods:

• Utilized acoustic droplet ejection for nanoliter-volume sample transfer in 1536-well plates.
• Developed the acoustic reverse phase protein array (acoustic RPPA) for multiplexable, low-cost immunodetection of native secreted proteins.
• Validated assay performance using the LOPAC1280 chemical library and a secreted bioluminescent reporter.

Main Results:

• Demonstrated comparable performance between acoustic RPPA and bioluminescent assays in identifying secretory modulators.
• Established a platform coupling extracellular protein quantification with intracellular and cytotoxicity analyses from single wells.
• Showcased proof-of-principle applications using human induced pluripotent stem cell-derived hepatocytes.

Conclusions:

• The developed platform offers a robust and multiplexed approach for drug discovery targeting secreted proteins.
• Acoustic RPPA is a versatile technology for analyzing native, endogenously secreted proteins in relevant model systems.
• This integrated platform facilitates comprehensive cellular analysis for identifying novel therapeutic strategies.