FRET에 기반한 Open Sandwich FluoroimmunoassayDevelopment of an Open Sandwich FluoroimmunoassayBased on FRET
- Other Titles
- Development of an Open Sandwich FluoroimmunoassayBased on FRET
- Authors
- Quande Wei; 이문권; 성기훈; 주재범; 이은규
- Issue Date
- Dec-2007
- Publisher
- 한국생물공학회
- Keywords
- FRET; sandwich assay; quantum dots; fluorescence labels; sandwich immunoassay; FRET; sandwich assay; quantum dots; fluorescence labels; sandwich immunoassay
- Citation
- KSBB Journal, v.22, no.6, pp 426 - 432
- Pages
- 7
- Indexed
- KCI
- Journal Title
- KSBB Journal
- Volume
- 22
- Number
- 6
- Start Page
- 426
- End Page
- 432
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/43979
- ISSN
- 1225-7117
- Abstract
- QDs을 기반으로 하는 OsFIA는 매우 빠르고 간단히 수행될 수 있다. 또한 이 분석법은 고체상의 담체나 결합/잔류 시약의 분리 등과 같은 여러 과정을 필요로 하지 않으며, 적은 양의 시약으로도 분석이 가능하다. 본 분석법은 높은 감도로 항원을 측정할 수 있으며, 일상적인 분석에도 쉽게 도입될 수 있을 것이다. 선형 범위 내에서 측정 가능한 receptor의 최소농도는 0.05 nM (2.65 ng/mL) 정도이다. 또한, 일반적으로 상용화된 항체를 가지고 수행이 가능하다. 이 OsFIA 분석법은 기존의 실험적 sandwich immunoassay의 효과적인 대안으로 제시된다.
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 Å. 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 > 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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