Droplet-free Digital ELISA法によるたんぱく質の高感度同時多項目検出

カテゴリ: 論文誌(論文単位)

グループ名: 【E】センサ・マイクロマシン部門

発行日: 2018/07/01

タイトル(英語): Highly Sensitive Multiplex Protein Detection by Droplet-free Digital ELISA

著者名: 赤間　健司（シスメックス（株）），白井　健太郎（シスメックス（株）），鈴木　誓吾（シスメックス（株））

著者名(英語): Kenji Akama (Sysmex Corporation), Kentaro Shirai (Sysmex Corporation), Seigo Suzuki (Sysmex Corporation)

キーワード: デジタルELISA，バイオセンサ，たんぱく質，高感度検出，多項目検出，チラミド　　Digital ELISA，biosensor，protein，high sensitive detection，multiplex detection，tyramide

要約(英語): Digital enzyme-linked immunosorbent assay (ELISA) is a single molecule counting technology and is one of the most sensitive immunoassay methods. The key aspect of this technology is to concentrate enzyme reaction products from a single target molecule in femtoliter droplets. On the other hand, we have developed a novel Digital ELISA that does not require droplets; instead, enzyme reaction products are concentrated using a tyramide signal amplification system. In this study, we present a method that enables the multiplex detection of proteins based on Droplet-free Digital ELISA. First, two types of paramagnetic beads immobilized with antibodies specific to different target proteins were incubated with a sample solution, and target proteins were captured on their specific beads. Then, these beads were labeled with horseradish peroxidase (HRP), and tyramide substrate reacted with HRP on beads, resulting in products of this reaction deposited on those beads. This signal amplification on beads makes it possible to count the number of labeled beads digitally. We used this approach to simultaneously detect IL-6 and HBs Ag with single molecule resolution. The obtained limit of detections were 0.1 pg/mL and 0.013 IU/mL, respectively. Our method has potential applications in simple in vitro diagnostic systems for simultaneous and high sensitive detection of protein biomarkers.

本誌: 電気学会論文誌E（センサ・マイクロマシン部門誌） Vol.138 No.7 （2018） 特集：第34回「センサ・マイクロマシンと応用システム」シンポジウム受賞論文

本誌掲載ページ: 270-274 p

原稿種別: 論文／日本語

電子版へのリンク: https://www.jstage.jst.go.jp/article/ieejsmas/138/7/138_270/_article/-char/ja/