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以一种纯天然产物——白藜芦醇(resveratrol,RST)作为辣根过氧化物酶(HRP)荧光底物运用于酶联荧光免疫传感体系.RST对HRP,H2O2的荧光响应性能优于传统HRP荧光底物,诸如对羟苯丙酸、AmplexRed和佳味醇.RST本身只有极弱的荧光,在HRP催化下可被H2O2氧化成二聚体产物,该二聚体在315nm的激发光下能发射波长为462nm的强荧光,并且反应体系的荧光强度增加值与HRP量在一定浓度范围内成线性相关.根据此关系和竞争型免疫定量原理,以日本血吸虫抗体(SjAb)为模型分析对象,建立了基于新HRP荧光底物的酶联荧光传感分析新方法.运用制备的传感装置测定SjAb的线性范围为1.5×10-6～7.3×10-4g/L,检出限为1.5×10-6g/L,测定浓度为5×10-6g/LSjAb的相对标准偏差为4.7%(n=10).RST的二聚体产物水溶性很低,通过该装置可将酶反应产物沉积在具Ag/SiO2纳米粒子的传感界面上,利用纳米Ag/SiO2对产物吸附富集作用,不仅解决了传统方法不便于测定低水溶性的产物的问题,而且提高了分析灵敏性.
A pure natural product, resveratrol (RST), was used as a fluorescent horseradish peroxidase (HRP) fluorescent substrate in the enzyme-linked immunosensor system. The fluorescence response of RST to HRP and H2O2 was excellent In traditional HRP fluorogenic substrates such as p-hydroxyphenylpropionate, Amplex Red, and good taste alcohol, RST itself has only very weak fluorescence and is oxidized by H2O2 to a dimeric product under HRP catalysis, which is excited at 315 nm Under the light, strong fluorescence with a wavelength of 462nm can be emitted, and the added value of the fluorescence intensity of the reaction system is linearly correlated with the amount of HRP within a certain concentration range.According to this relationship and the competitive immunological quantification principle, the SjAb antibody was used as a model And a new method of enzyme-linked fluorescence detection based on the new HRP fluorogenic substrate was established.Using the prepared sensor, the linear range of SjAb was 1.5 × 10-6 ~ 7.3 × 10-4g / L, the detection limit Was 1.5 × 10-6g / L and the relative standard deviation was 4.7% (n = 10) at a concentration of 5 × 10-6g / LSjAb. The dimer product of RST was very poorly water-soluble, and the enzyme reaction The product was deposited on a sensing interface with Ag / SiO2 nanoparticles, and the product was adsorbed and enriched using nano-Ag / SiO2 , Not only solved the problem of the conventional measurement method is not easy to product low water solubility, but also improve analytical sensitivity.