Development of detection method for multiple disease-causative proteins using dried blood spots

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K07877
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 52050:Embryonic medicine and pediatrics-related
• Research Institution: Kazusa DNA Research Institute
• Principal Investigator: Nakajima Daisuke 公益財団法人かずさDNA研究所, ゲノム事業推進部, 研究員 (30370935)
• Co-Investigator(Kenkyū-buntansha): 川島 祐介 公益財団法人かずさDNA研究所, ゲノム事業推進部, ユニット長 (30588124)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: 乾燥ろ紙血 / 新生児スクリーニング / プロテオーム解析

Outline of Final Research Achievements

Protein extraction from dried blood spots using the sodium carbonate precipitation method was investigated and a sample preparation method suitable for deep proteome analysis was established. We have also established LC-MS conditions specifically for the deep analysis of proteins extracted from dried blood spots. As a result, more than 3,000 proteins, including more than 1,000 disease-causing proteins, could be identified and quantified from dried blood spots by one shot LC-MS analysis. Furthermore, we have confirmed that 38 stable isotope-labelled peptides required for the quantification of disease-causing proteins can be produced cheaply and simultaneously by combining a cell-free expression system from E. coli with a multiplex co-expression system.

Free Research Field

胎児医学および小児育成学関連

Academic Significance and Societal Importance of the Research Achievements

新生児スクリーニングは先制医療としてその重要性を増している。しかしながらターゲットとなる代謝物や酵素をそれぞれ個別の方法で検出しているため検査項目が増えるにつれコストや時間が嵩むという問題を抱えている。本研究で確立した炭酸ナトリウム沈殿法を用いた乾燥ろ紙血由来の蛋白質濃縮法を用いれば1,000以上の疾患原因蛋白質を含む3,000以上の蛋白質を同定・定量することができる。これにより安価で迅速かつ網羅的な新生児スクリーニング法の開発が進むと考えられる。