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基于树枝状聚合物-卟啉双发光分子标记的小麦凝集素亲和吸附固体基质室温磷光法测定痕量葡萄糖及人类疾病预测

Determination of trace glucose and forecast of human diseases by affinity adsorption solid substrate-room temperature phosphorimetry based on triticum vulgaris lectin labeled with dendrimers-porphyrin dual luminescence molecule.

作者信息

Liu Jia-Ming, Liu Zhen-Bo, Zhu Guo-Hui, Li Xue-Lin, Huang Xiao-Mei, Li Fei-Ming, Shi Xiu-Mei, Zeng Li-Qing

机构信息

Department of Chemistry and Environmental Science, Zhangzhou Normal College, Zhangzhou 363000, PR China.

出版信息

Talanta. 2008 Jan 15;74(4):625-31. doi: 10.1016/j.talanta.2007.06.040. Epub 2007 Jul 5.

DOI:10.1016/j.talanta.2007.06.040
PMID:18371685
Abstract

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.

摘要

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。

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