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KMID : 0380020070220060426
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Korean Journal of Biotechnology and Bioengineering
2007 Volume.22 No. 6 p.426 ~ p.432
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Development of an Open Sandwich Fluoroimmunoassay Based on FRET
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Wei Quan-De
Lee Mon-Kwon
Seong Gi-Hun
Choo Jae-Bum
Lee Eun-Kyu
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Abstract
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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- (ER-) antigen was incubated with QD-labeled anti-ER- monoclonal antibody and AF (Alexa Fluoro)-labeled anti-ER polyclonal antibody for 30 minutes, followed by FRET measurement. The dye separation distance was estimated to be between . The present method is rapid, simple and highly sensitive, and 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 ( > 0.990). In addition, the assay was performed with commercial antibodies. This assay provides a convenient alternative to conventional, laborious sandwich immunoassays.
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KEYWORD
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FRET, sandwich assay, quantum dots, fluorescence labels, sandwich immunoassay
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