CHAPTER 11: Discovery and Validation Case Studies, Recommendations: Discovery and Development of Multimarker Panels for Improved Prediction of Near‐Term Myocardial Infarction

This report is a case study of the complex process of designing and performing a biomarker discovery study, validating its preliminary findings, and translating these into a clinical utility that would be suitable for the requirements of FDA clearance. The process is illustrated on the development of a multianalyte (protein) panel that can significantly enhance the prediction of the risk of near‐term myocardial infarction (MI) over traditional risk factors. The utilized clinical material is a subset of the Copenhagen General Population Study consisting of a matched set 252 cases, defined as fatal or nonfatal MI within four years of blood collection, and 499 controls. For discovery, we utilized high‐resolution mass‐spectrometric profiling of pooled samples complemented with multiplexed immunoassays. Discoveries were validated by using multiple reaction monitoring (MRM) liquid chromatography‐mass spectrometry (LC‐MS). It was demonstrated that a six‐protein panel comprised of analytes measured by MRM and immunoassays could significantly improve risk prediction over that provided by traditional risk factors alone. The path of developing a “submission package” for FDA clearance is also discussed.