The problem:
Assay has low s/b (?:1), excessive variability, reagents are unstable, readout is not robot-friendly e.g. filter binding.
- need to improve assay robustness
- need to reformat readout to introduce new technologies.
Multiple variables are involved
- time, temperature, order of addition
- buffer composition, pH
- reagent concentrations
- readout conditions e.g. SPA, fluorescence technologies
Need to optimize conditions for reagent scale-up
How do you find the best combination of variables that gives a robust, robot-friendly signal?
Consider an assay with only 2 variables -salt and pH.