Li Zhiming, Zhu Guohui, Liu Jiaming, Lu Qiaomei, Yang Minlan, Wu Hong, Shi Xiumei, Chen Xinhua
Department of Food and Biological Engineering, Zhangzhou Institute of Technology, Zhangzhou 363000, PR China.
Spectrochim Acta A Mol Biomol Spectrosc. 2007 Aug;67(5):1153-60. doi: 10.1016/j.saa.2006.11.041. Epub 2006 Dec 15.
A new phosphorescence labeling reagent Triton-100X-4.0G-D (4.0G-D refers to 4.0-generation dendrimers) was found. Quantitative specific affinity adsorption (AA) reaction between Triton-100X-4.0G-D-WGA and glucose (G) was carried out on the surface of nitrocellulose membrane (NCM), and the DeltaI(p) of the product of AA reaction was linear correlation to the content of G. Based on the facts above, a new method for the determination of trace G was established by WGA labeled with Triton-100X-4.0G-D affinity adsorption solid substrate room temperature phosphorimetry (Triton-100X-4.0G-D-WGA-AA-SS-RTP). This research showed that AA-SS-RTP for either direct method or sandwich method could combine very well the characteristics of both the high sensitivity of SS-RTP and the specificity of the AA reaction. Detection limits (LD) were 0.24 fg spot(-1) for direct method and 0.18 fg spot(-1) for sandwich method, indicating both of them were of high sensitivity. The method has been applied to the determination of the content of G in human serum, and the results were coincided with those obtained by glucose oxidize enzyme method. It can also be applied to forecast accurately some human diseases, such as primary hepatic carcinoma, cirrhosis, acute and chronic hepatitis, transfer hepatocellular, etc. Meanwhile, the mechanism for the determination of G with AA-SS-RTP was discussed.
发现了一种新型磷光标记试剂Triton-100X-4.0G-D(4.0G-D指第四代树枝状大分子)。在硝酸纤维素膜(NCM)表面进行了Triton-100X-4.0G-D-WGA与葡萄糖(G)之间的定量特异性亲和吸附(AA)反应,AA反应产物的ΔI(p)与G的含量呈线性相关。基于上述事实,建立了一种用Triton-100X-4.0G-D标记的WGA亲和吸附固体基质室温磷光法(Triton-100X-4.0G-D-WGA-AA-SS-RTP)测定痕量G的新方法。本研究表明,无论是直接法还是夹心法的AA-SS-RTP都能很好地结合SS-RTP的高灵敏度和AA反应的特异性这两个特点。直接法的检测限(LD)为0.24 fg spot(-1),夹心法为0.18 fg spot(-1),表明两者均具有高灵敏度。该方法已应用于人血清中G含量的测定,结果与葡萄糖氧化酶法一致。它还可用于准确预测一些人类疾病,如原发性肝癌、肝硬化、急慢性肝炎、转移性肝细胞癌等。同时,对AA-SS-RTP测定G的机理进行了探讨。