Development of an Open Sandwich Fluoroimmunoassay Based on FRET
2007
Wei, Quande (Sun Yat-sen University, Guangzhou, China) | Lee, M.K. (Hanyang University, Ansan, Republic of Korea) | Seong, G.H. (Hanyang University, Ansan, Republic of Korea) | Choo, J.B. (Hanyang University, Ansan, Republic of Korea) | Lee, E.K. (Hanyang University, Ansan, Republic of Korea), E-mail: [email protected]
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 AF (Alexa Fluoro)-labeled anti-ER polyclonal antibody for 30 minutes, followed by FRET measurement. The dye separation distance was estimated to be between 80~90 a with ring above. 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 (R² greater than 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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