title: Development of an open sandwich fluoroimmunoassay based on fluorescence resonance energy transfer.
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 A. 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.
authors.affiliation: Department of Pathogenic Biology, Sun Yat-sen University, Guangzhou 530080, People's Republic of China.
authors.forename: Quande
authors.initials: Q
authors.lastname: Wei
authors.forename: Moonkwon
authors.initials: M
authors.lastname: Lee
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
authors.forename: Eun Kyu
authors.initials: EK
authors.lastname: Lee
authors.forename: Gi Hun
authors.initials: GH
authors.lastname: Seong
authors.forename: Jaebum
authors.initials: J
authors.lastname: Choo
authors.forename: Yong Woo
authors.initials: YW
authors.lastname: Cho
mesh_terms: D000911:Antibodies, Monoclonal; D047629:Estrogen Receptor beta / Q000032:analysis* / Q000276:immunology; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes; D015200:Fluoroimmunoassay / Q000379:methods*; D008956:Models, Chemical; D045663:Quantum Dots; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D047629:Estrogen Receptor beta; D005456:Fluorescent Dyes