Identify biomarker

Comes from exploratory or focused analysis
Assay metrics optimized
Analytical cut-offs defined
Fit for intended purpose

Prospective validation

Integrated into prospective study
Powered for secondary analysis using validated fixed discriminants

Ready for application

Use as selection, stratification, endpoint
Stratification and selection require CLIA lab

Retrospective validation

Correlation with clinical variable vetted
Powered independent dataset
Uses defined fixed discriminants

End