Development of an open sandwich fluoroimmunoassay based on fluorescence resonance energy transfer
- 作者：Quande Wei ; Moonkwon Lee ; Xinbing Yu ; Eun Kyu Lee ; Gi Hun Seong ; Jaebum Choo and Yong Woo Cho
- 刊名：Analytical Biochemistry
- 出版年：2006
- 期刊代码：93_00032697
- 类别：imm
- 出版时间：1 November 2006
- 卷：358
- 期：1
- 页码：31-37
- 文件大小：359 K
摘要
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 β (ER-β) antigen was incubated with QD-labeled anti-ER-β 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 Å. 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.