Augustin Christine M, Oswald Bernhard, Wolfbeis Otto S
Institute of Analytical Chemistry, University of Regensburg, Regensburg, D-93040, Germany.
Anal Biochem. 2002 Jun 15;305(2):166-72. doi: 10.1006/abio.2002.5633.
A novel immunosystem is described that exploits the effect of luminescence energy transfer from a luminescently labeled antigen to a fluorescent antibody. A luminescent ruthenium-ligand complex (D-455) with absorption/emission maxima at 456/639 nm, respectively, was employed as the donor label, and a squaraine-type cyanine label (636/655 nm), as the fluorescent acceptor label. Specifically, the system human serum albumin (HSA)/anti-HSA was studied. HSA was labeled with the donor dye D-455, and anti-HSA was labeled with the acceptor dye A-631. On formation of the antigen-antibody complex, energy transfer occurs. The radiationless energy transfer affects both the decay time of D-455 and the intensities of the emissions of both D-455 and A-631. The decay time of around 500 ns of D-455 allows frequency-domain measurements in the low kilohertz range and therefore can be based on the use of conventional optoelectronics. This also suggests gated measurements to be performed. The major difference from existing HSA immunosystems is the use of a slow decaying ruthenium-ligand complex as the donor and of a long-wave emitting cyanine acceptor dye having a high quantum yield and a decay kinetics that is governed by the rate of energy transfer from the slow decaying donor.
描述了一种新型免疫系统，该系统利用从发光标记抗原到荧光抗体的发光能量转移效应。分别在456/639 nm处具有最大吸收/发射峰的发光钌-配体复合物（D-455）用作供体标记，方酸菁型花菁标记（636/655 nm）用作荧光受体标记。具体而言，研究了人血清白蛋白（HSA）/抗HSA系统。HSA用供体染料D-455标记，抗HSA用受体染料A-631标记。抗原-抗体复合物形成时，会发生能量转移。无辐射能量转移会影响D-455的衰减时间以及D-455和A-631的发射强度。D-455约500 ns的衰减时间允许在低千赫兹范围内进行频域测量，因此可以基于传统光电子学的使用。这也表明可以进行门控测量。与现有的HSA免疫系统的主要区别在于使用缓慢衰减的钌-配体复合物作为供体，以及使用具有高量子产率和由缓慢衰减供体的能量转移速率控制的衰减动力学的长波发射花菁受体染料。
