Rapid detection of two-protein interaction with a single fluorophore by using a microfluidic device

Chao Kai Chou, Nan Jing, Hirohito Yamaguchi, Pei Hsiang Tsou, Heng Huan Lee, Chun Te Chen, Ying Nai Wang, Sungmin Hong, Chin Su, Jun Kameoka*, Mien Chie Hung

*この研究の対応する著者

研究成果: Article査読

9 被引用数 (Scopus)

抄録

We have developed a microfluidics based platform and methodology named MAPS (microfluidic system for analyzing proteins in single complex) for detecting two protein interactions rapidly using a single fluorophore. Target proteins were labelled with Quantum dot 525 (QD525) via specific polyclonal antibodies, and were transported through the microfluidic channel subsequently, where the 375 nm excitation laser light was focused to form a detection volume. Photon bursts from target proteins passing through the detection volume were recorded and their photon burst histograms were plotted which demonstrated roughly the specific protein interaction ratio based on their population and statistical behavior. As a proof of concept, Src/STAT3 protein complex interaction ratios with and without EGF stimulation were obtained by MAPS within 1 h and the results were well matched with the one obtained by the conventional immunoprecipitation/Western blot (IP/WB).

本文言語English
ページ(範囲)2907-2912
ページ数6
ジャーナルAnalyst
135
11
DOI
出版ステータスPublished - 2010 11月
外部発表はい

ASJC Scopus subject areas

  • 分析化学
  • 生化学
  • 環境化学
  • 分光学
  • 電気化学

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