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迈向新型新生儿筛查方法的发展：使用低成本铁粉通过机器人流程对干血斑进行深入非靶向蛋白质组分析。

Toward the Development of a Novel Newborn Screening Modality: In-Depth Nontargeted Proteome Analysis of Dried Blood Spots with a Robotic Pipeline Using Low-Cost Iron Powders.

作者信息

Nakajima Daisuke, Ishikawa Masaki, Konno Ryo, Okuda Yusei, Sasai Hideo, Ohara Osamu, Kawashima Yusuke

机构信息

Department of Applied Genomics, Kazusa DNA Research Institute, 2-5-23 Kazusa Kamatari, Kisarazu, Chiba 292-0818, Japan.

出版信息

Anal Chem. 2025 Aug 26;97(33):17992-18000. doi: 10.1021/acs.analchem.5c01720. Epub 2025 Aug 11.

DOI:10.1021/acs.analchem.5c01720
PMID:40790917
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12392252/
Abstract

We developed a simple protein extraction method for dried blood spots (DBS) that potentially meets the throughput required for newborn screening (NBS) and optimizes nontargeted proteomic analysis in combination with liquid chromatography coupled mass spectrometry in the data-independent-acquisition mode (DIA-LC-MS/MS). The developed pipeline, termed on-targeted nalysis of on-specifically BS-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 45 samples/day with minimal compromise in protein coverage depth. The results imply that this pipeline can open new venues for conducting NBS using nontargeted quantitative proteome profiling, which has been a missing modality in NBS.

摘要

我们开发了一种用于干血斑（DBS）的简单蛋白质提取方法，该方法有可能满足新生儿筛查（NBS）所需的通量，并结合液相色谱-串联质谱的数据非依赖采集模式（DIA-LC-MS/MS）优化非靶向蛋白质组分析。所开发的流程称为特异性吸附于干血斑的蛋白质的靶向分析（NANDA），成功解决了以下三个挑战：（1）使用低成本铁粉通过机器人系统并行处理96个3.2毫米的DBS冲孔；（2）使用DIA-LC-MS/MS鉴定出5000多种蛋白质；（3）将DIA-LC-MS/MS通量提高到每天45个样本，同时在蛋白质覆盖深度上的折衷最小。结果表明，该流程可为利用非靶向定量蛋白质组谱进行新生儿筛查开辟新途径，而这在新生儿筛查中一直是一种缺失的方式。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/d35c8fd9a0b9/ac5c01720_0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/6492cb960b92/ac5c01720_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/aef9e030ce5b/ac5c01720_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/5cc4f967e426/ac5c01720_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/62d9dd18c5b0/ac5c01720_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/5263ab273ea8/ac5c01720_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/ceecc6ec444c/ac5c01720_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/d35c8fd9a0b9/ac5c01720_0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/6492cb960b92/ac5c01720_0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/aef9e030ce5b/ac5c01720_0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/5cc4f967e426/ac5c01720_0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/62d9dd18c5b0/ac5c01720_0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/5263ab273ea8/ac5c01720_0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/ceecc6ec444c/ac5c01720_0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/61b4/12392252/d35c8fd9a0b9/ac5c01720_0007.jpg

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