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Development of an open sandwich fluoroimmunoassay based on fluorescence resonance energy transfer
- Authors
- Wei, Quande; Lee, Moonkwon; Yu, Xinbing; Lee, Eun Kyu; Seong, Gi Hun; Choo, Jaebum; Cho, Yong Woo
- Issue Date
- Nov-2006
- Publisher
- ACADEMIC PRESS INC ELSEVIER SCIENCE
- Keywords
- FRET; sandwich assay; quantum dots; fluorescence labels; sandwich immunoassays
- Citation
- ANALYTICAL BIOCHEMISTRY, v.358, no.1, pp.31 - 37
- Indexed
- SCIE
SCOPUS
- Journal Title
- ANALYTICAL BIOCHEMISTRY
- Volume
- 358
- Number
- 1
- Start Page
- 31
- End Page
- 37
- ISSN
- 0003-2697
- Abstract
- We have developed a sensitive, one-step, homogeneous open sandwich fluoroimmunoassay (OsFIA) based on fluorescence resonance energy transfer (FRET) and luminescent semiconductor quantum dots (QDs). In this FRET assay, estrogen receptor beta (ER-beta) antigen was incubated with QD-labeled anti-ER-beta monoclonal antibody and Alexa Fluor (AF)-labeled anti-ER polyclonal antibody for 30 min, followed by FRET measurement. The dye separation distance was estimated between 80 and 90 angstrom. The current method is rapid, simple, and highly sensitive, and it did not require the bound/free reagent separation steps and solid-phase carriers. A concentration as low as 0.05 nM (2.65 ng/ml) receptor was detected with linearity. In addition, the assay was performed with commercial antibodies. This assay provides a convenient alternative to conventional, laborious sandwich immunoassays. (c) 2006 Elsevier Inc. All rights reserved.
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Related Researcher
- Cho, Yong Woo
- ERICA 공학대학 (DEPARTMENT OF MATERIALS SCIENCE AND CHEMICAL ENGINEERING)