Assay robustness is improved through optimization

• Robustness is equivalent to high sensitivity to controlled factors plus low sensitivity to uncontrolled variables

  • controlled variables are those you want to influence e.g. ligand or substrate concentration
  • uncontrolled variables are ‘error’ or ‘incidental’ factors e.g. pipetting variance (volume), temperature, DMSO, pH etc.

• More specifically, a robust assay minimizes the slope of the response surface for uncontrolled variables:

noise

no noise

volume

s/b

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