Section7:Requirements for Assay Proof of Concept for Cell Based Assays

From Assay Guidance Wiki

1. Receptor of interest has been expressed in a suitable cell line and functional expression has been verified. Receptor expression has been verified by RT-PCR or Western blot.
2. Sample preparation has passed QC guidelines above.
3. Preliminary source for all reagents has been identified.
4. Early passage stable cell lines are available and free of mycoplasma contamination.
5. Transient transfection assays: appropriate cell line and transfection procedure demonstrated. Plasmids available and passes QC sequence and restriction mapping criteria (above).
6. Cell culture details are available as written SOP. This includes the number of cells (not dilution eg 1/10) used for passage, passage frequency, limit passage numbers for an assay, activity stability as a function of cell passage and density, and optimum cell density for target activity.
7. Biological activity (>90%) is target specific as demonstrated by transfection controls, comparison to parental cell lines, pharmacology, and/or tool compound activity.
8. Assay signal is dependant upon amount of cells present.
9. Assays enabled in DHT: Preliminary data showing a saturable activator and/or inhibitor response with sufficient signal window using a QB-supported assay format.