Automation of a Novel Cell-Based ELISA for Cell Signaling Pathway Analysis

In addition to offering simplicity, speed, and flexibility, the technology is a highly sensitive technique that lends itself to automation at multiple levels including seeding cells in microplates; cell media exchanges; serial dilution and transfer steps; assay reagent additions; microplate washing; and, signal detection. Assay performance studies were conducted using lysate kit controls to verify optimal automation performance for S/B, Z’, and intra- and inter-assay precision. A ‘one well one day’ cell based assay protocol was developed and used for pharmacology assessment for agonist/antagonist analysis of the PI-3/AKT/mTOR signaling pathway. Procedures demonstrated here are well suited for validating automation of model system protocol development.