4.0代树枝状分子-卟啉标记伴刀豆凝集素亲和吸附固体基质室温磷光法测定人血清中甲胎蛋白异质体

Determination of Alpha-fetoprotein Variant in Human Serum by Affinity Adsorption Solid Substrate-room Temperature Phosphorimetry Based on Concanavalin A Labeled with 4.0 Generation Dendrimer-porphyrin

中文关键词:  甲胎蛋白异质体,树枝状分子-卟啉,标记,伴刀豆凝集素,亲和吸附固体基质室温磷光法

英文关键词:alpha-fetoprotein variant,dendrimer-porphyrin,label,concanavalin A,affinity adsorption solid substrate-room temperature phosphorimetry

中文摘要:

      提出一种4.0代树枝状分子(4.0G-D)-卟啉(P)双发光分子新的磷光标记试剂（4.0G-D-P）.基于4.0G-D-P标记伴刀豆凝集素(Con A）的产物（4.0G-D-P-Con A）能在聚酰胺素膜(PAM)上发射强而稳定的室温磷光（RTP），而且该标记产物能与甲胎蛋白异质体（AFP-V）发生特异性的亲和吸附（AA）反应，其反应产物保持了4.0G-D-P双发光分子RTP的优良特性，且ΔI_p值与AFP-V的含量呈线性关系，加入Tween-80可以提高ΔI_p值，据此建立了Tween-80-4.0G-D-P双发光分子标记Con A亲和吸附固体基质室温磷光法（AA-SS-RTP）测定人血清中AFP-V的新方法.直接法的检出限为0.31 pg/mL（4.0G-D）和0.43 pg/mL（P），灵敏度较高.无论用4.0G-D或P的激发/发射波长测定人血清中AFP-V的含量，结果与酶联免疫法（ELISA）相吻合.

英文摘要:

      The 4.0 generation dendrimer (4.0G-D)-porphyrin (P) dual luminescence molecule (4.0G-D-P) is brought forward as a new phosphorescence label reagent. The product (4.0G-D-P-Con A) of 4.0G-D-P labeling concanavalin A (Con A) can emit strong and stable room temperature phosphorescence (RTP) signal on the surface of polyamide membrane (PAM)，and it was also found that there is a specific affinity adsorptive (AA) reaction between 4.0G-D-P-Con A and alpha-fetoprotein variant (AFP-V). The product of the reaction can keep very well the RTP characteristics of 4.0G-D-P, and the ΔI_pwas linear correlation to the content of AFP-V. When adding Tween-80, the ΔI_pwas increase. According to the facts above, a new method of affinity adsorption solid substrate-room temperature phosphorimetry for determination of AFP-V in human serum was established basing on Con A labeled with tween-80-4.0G-D-P dual luminescence molecule. The detection limits of this method were 0.31 pg/mL with 4.0G-D and 0.43 pg/mL with P for direct assay. Determination of AFP-V in human serum with 4.0G-D or P was as well as enzyme-linked immunosorbent assay (ELISA), the sensitivity was obviously improved and the applicability was wider.