Use of a long lifetime Re(I) complex in fluorescence polarization immunoassays of high-molecular weight analytes
Use of a long lifetime Re(I) complex in fluorescence polarization immunoassays of high-molecular weight analytes
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DOI：
10.1021/ac970827k
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发表时间：
1998-02-01
影响因子：
7.4
通讯作者：
Lakowicz, JR
中科院分区：
文献类型：
--
作者：
Guo, XQ;Castellano, FN;Lakowicz, JR
本文描述了一类新的基于Re（I）金属-配体络合物发光的荧光偏振免疫分析方法。Re（I）金属-配体络合物具有极高的光稳定性和有用的物理化学性质，包括长寿命、高量子产率和在没有旋转扩散的情况下的高发射偏振。[Re（bcp）（CO）（3）（4-COOHPy）]（ClO 4），其中bcp是2，9-二甲基-4，7-二苯基-1，10-菲咯啉，4-COOHPy是异烟酸，已被评价用于高分子量抗原的荧光偏振免疫测定（FPI）。该Re（I）络合物（Re）在无旋转扩散的情况下显示出高度偏振的发射（最大各向异性接近0.3），并且当在含氧水溶液中与人血清白蛋白（HSA）结合时显示出长的平均寿命（2.7 μ s）。这些实验结果与理论预测相结合，证明了这种Re（I）金属-配体络合物作为高分子量分析物（10（5）-10（8）Da）的FPI的发光探针的潜力。
We describe a new class of fluorescence polarization immunoassays based on the luminescence from a Re(I) metal-ligand complex, Re(I) complexes are extremely photostable and possess useful photophysical properties including long lifetimes, high quantum yields, and high emission polarization in the absence of rotational diffusion, In the present study, a conjugatable, highly luminescent Re(I) metal-ligand complex, [Re(bcp)(CO)(3)(4-COOHPy)](ClO4), where bcp is 2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline and 4-COOHPy is isonicotinic acid, has been evaluated for use in fluorescence polarization immunoassays (FPIs) with high-molecular-weight antigens. This Re(I) complex (Re) displays highly polarized emission (with a maximum anisotropy near 0.3) in the absence of rotational diffusion and a long average lifetime (2.7 mu s) when bound to human serum albumin (HSA) in oxygenated aqueous solution, The emission polarization of the Re-HSA conjugate is sensitive to the binding of anti-HSA, resulting in a significant increase in anisotropy. The labeled HSA was also used in a competition immunoassay where unlabeled HSA was also used as an antigen, These experimental results, combined with theoretical predictions, demonstrate the potential of this Re(I) metal-ligand complex as a luminescence probe in FPIs of high-molecular-weight analytes (10(5)-10(8) Da).