Detecting Secretory Proteins by Acoustic Droplet Ejection in Multiplexed High-Throughput Applications
| Autor: | Michael J. Iannotti, Ryan MacArthur, Richard E. Jones, Sam Michael, James Inglese, Dingyin Tao, Ilyas Singeç |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Quantitative proteomics Induced Pluripotent Stem Cells Biomedical Technology Protein Array Analysis 01 natural sciences Biochemistry Article Antibodies Chemical library Cell Line Small Molecule Libraries 03 medical and health sciences chemistry.chemical_compound Drug Discovery Humans Induced pluripotent stem cell Acoustic droplet ejection 010405 organic chemistry Chemistry Drug discovery Reverse phase protein lysate microarray General Medicine Acoustics 0104 chemical sciences Cell biology High-Throughput Screening Assays 030104 developmental biology Secretory protein Proteome Hepatocytes Molecular Medicine |
| Zdroj: | ACS chemical biology. 14(3) |
| ISSN: | 1554-8937 |
| Popis: | Nearly one third of the encoded proteome is comprised of secretory proteins that enable communication between cells and organ systems, playing a ubiquitous role in human health and disease. High-throughput detection of secreted proteins would enhance efforts to identify therapies for secretion-related diseases. Using the Z mutant of alpha-1 antitrypsin as a human secretory model, we have developed 1536-well high-throughput screening assays that utilize acoustic droplet ejection to transfer nanoliter volumes of sample for protein quantification. Among them, the acoustic reverse phase protein array (acoustic RPPA) is a multiplexable, low-cost immunodetection technology for native, endogenously secreted proteins from physiologically relevant model systems like stem cells that is compatible with plate-based instrumentation. Parallel assay profiling with the LOPAC(1280) chemical library validated performance and orthogonality between a secreted bioluminescent reporter and acoustic RPPA method by consistently identifying secretory modulators with comparable concentration response relationships. Here, we introduce a robust, multiplexed drug discovery platform coupling extracellular protein quantification by acoustic RPPA with intracellular and cytotoxicity analyses from single wells, demonstrating proof-of-principle applications for human induced pluripotent stem cell-derived hepatocytes. |
| Databáze: | OpenAIRE |
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