A novel newborn screening modality: Non-targeted proteome analysis using low-cost iron powders
Nakajima, D.; Ishikawa, M.; Konno, R.; Sasai, H.; Ohara, O.; Kawashima, Y.
Show abstract
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 Non-targeted Analysis of Non-specifically DBS-Absorbed 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.
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