A novel newborn screening modality: Non-targeted proteome analysis using low-cost iron powders

In this study, we developed a simple protein extraction method for dried blood spots (DBS) that potentially meets the throughput required for newborn screening (NBS) and optimizes non-targeted proteomic analysis in combination with liquid chromatography coupled mass spectrometry in the data-independent-acquisition mode (DIA-LC-MS/MS). The developed pipeline, termed N on-targeted A nalysis of N on-specifically D BS- A bsorbed proteins (NANDA), successfully addressed the following three challenges: (1) processing of 96 3.2-mm DBS punches in parallel using low-cost iron powders with a robotic system, (2) identifying more than 5,000 proteins using DIA-LC-MS/MS, and (3) improving DIA-LC-MS/MS throughput to 40 samples/day with minimal compromise in protein coverage depth. The results imply that this pipeline can open new venues for conducting NBS using non-targeted quantitative proteome profiling, which has been a currently missing modality in NBS.