Liu Jia-Ming, Liu Zhen-Bo, Zhu Guo-Hui, Li Xue-Lin, Huang Xiao-Mei, Li Fei-Ming, Shi Xiu-Mei, Zeng Li-Qing
Department of Chemistry and Environmental Science, Zhangzhou Normal College, Zhangzhou 363000, PR China.
Talanta. 2008 Jan 15;74(4):625-31. doi: 10.1016/j.talanta.2007.06.040. Epub 2007 Jul 5.
In this paper, 3.5-generation polyamidoamine dendrimers (3.5G-D)-porphyrin (P) dual luminescence molecule (3.5G-D-P) was developed as a new phosphorescence-labeling reagent. Meanwhile, the room temperature phosphorescence (RTP) characteristics of 3.5G-D-P and its product of labeling triticum vulgaris lectin (WGA) on the surface of polyamide membrane (PAM) were studied. Results showed that in the presence of heavy atom perturber LiAc, 3.5G-D and P of 3.5G-D-P molecule could emit strong and stable RTP on the PAM. And the Tween-80 would spike thoroughly the phosphorescence signal of 3.5G-D and P; moreover, specific affinity absorption (AA) reaction between the products (Tween-80-3.5G-D-P-WGA) of WGA labeled with Tween-80-3.5G-D-P and glucose (G) was carried out. The products of the AA reaction could keep good RTP characteristics of 3.5G-D and P dual luminescence molecule, and the DeltaI(p) was linear correlation to the content of G. According to the facts above, a new method of affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) for the determination of trace G was established, basing on WGA labeled with Tween-80-3.5G-D-P dual luminescence molecule. The detection limit of this method was 0.13fgspot(-1) (1.7x10(-12)moll(-1), 3.5G-D) and 0.14fgspot(-1) (2.2x10(-12)moll(-1), P). Determination of G in human serum using excitation/emission wavelength of either 3.5G-D or P, the result was coincided with enzyme-linked immunosorbent assay (ELISA). Not only the sensitivity and accuracy of this method were higher, but also the flexibility of AA-SS-RTP was obviously improved and the applicability was wider.
本文研制了一种新型磷光标记试剂——3.5代聚酰胺 - 胺树枝状大分子(3.5G - D) - 卟啉(P)双发光分子(3.5G - D - P)。同时,研究了3.5G - D - P及其标记小麦胚凝集素(WGA)的产物在聚酰胺膜(PAM)表面的室温磷光(RTP)特性。结果表明,在重原子微扰剂LiAc存在下,3.5G - D - P分子中的3.5G - D和P在PAM上能发射强而稳定的RTP。吐温 - 80会完全猝灭3.5G - D和P的磷光信号;此外,进行了吐温 - 80 - 3.5G - D - P标记的WGA产物(吐温 - 80 - 3.5G - D - P - WGA)与葡萄糖(G)之间的特异性亲和吸附(AA)反应。AA反应产物能保持3.5G - D和P双发光分子良好的RTP特性,且ΔI(p)与G的含量呈线性相关。基于上述事实,建立了一种基于吐温 - 80 - 3.5G - D - P双发光分子标记WGA的痕量G测定的亲和吸附固体基质 - 室温磷光法(AA - SS - RTP)。该方法的检测限为0.13 fgspot(-1)(1.7×10(-12) moll(-1),3.5G - D)和0.14 fgspot(-1)(2.2×10(-12) moll(-1),P)。采用3.5G - D或P的激发/发射波长测定人血清中的G,结果与酶联免疫吸附测定(ELISA)相符。该方法不仅灵敏度和准确度更高,而且AA - SS - RTP的灵活性明显提高,适用性更广。