Bazin H, Trinquet E, Mathis G
CIS bio international, Bagnols sur Céze, France.
J Biotechnol. 2002 Jan;82(3):233-50. doi: 10.1016/s1389-0352(01)00040-x.
Fluorescence resonance energy transfer (FRET) in association with a time-resolved fluorescence mode of detection was used to design a new homogeneous technology suitable to monitor biomolecular interactions. A lanthanide cryptate characterised by a long lived fluorescence emission was used as donor and a cross-linked allophycocyanine was used as acceptor. This new donor/acceptor pair displayed an exceptionally large Forster radius of 9 nm. This allowed to build up a set of labelling strategies to probe the interactions between biomolecules with an emphasis on fully indirect cassette formats particularly suitable for high throughput screening applications. Herein we describe the basics of the technology, review the latest applications to the study of molecular interactions involved in cells and new oligonucleotides based assays.
荧光共振能量转移(FRET)与时间分辨荧光检测模式相结合,用于设计一种适用于监测生物分子相互作用的新型均相技术。以长寿命荧光发射为特征的镧系穴合物用作供体,交联别藻蓝蛋白用作受体。这种新的供体/受体对显示出9nm的超大福斯特半径。这使得能够建立一套标记策略,以探测生物分子之间的相互作用,重点是特别适用于高通量筛选应用的完全间接盒式格式。在此,我们描述了该技术的基本原理,回顾了其在细胞中涉及的分子相互作用研究以及基于新寡核苷酸的检测方法中的最新应用。