In a publication in Clinical Proteomics a novel orthogonal strategy based on affinity proteomics and mass spectrometry was used for analytical validation of biomarkers in longitudinally collected serum samples from Duchenne muscular dystrophy (DMD) patients.

DMD is a fatal, incurable progressive neuromuscular disease, and even if discovery of protein biomarker candidates to aid development of novel therapies has been successful none of these candidates has yet been validated and translated into clinical use. This is partly attributed to biomarker discovery studies usually relying on high-throughput methods and relative quantification, while absolute quantification of biomarkers is required in clinical practice.

In this study quantification of the biomarkers using parallel reaction monitoring mass spectrometry (PRM-MS) was combined with sandwich immunoassays to simultaneously provide both an analytical validation of the biomarker concentration variation over time and a cross-validation of the proteomics methods. The results show that five out of ten biomarker candidates previously identified by affinity-based proteomics methods and found to be correlating with disease progression or severity, could be confirmed as associated with DMD using the mass spectrometry-based method.

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