基于表面增强拉曼光谱的碱性磷酸酶活性检测方法研究  

Detection of Alkaline Phosphatase Activity by using Surface-Enhanced Raman Spectroscopy

作　　者：江蕾 甘振飞 陈华英 常帅 李家宾 李大伟[1] JIANG Lei;GAN Zhenfei;CHEN Huaying;CHANG Shuai;LI Jiabin;LI Dawei(Key Laboratory for Advanced Materials,School of Chemistry and Molecular Engineering,East China University of Science and Technology,Shanghai 200237,P.R.China)

机构地区：[1]华东理工大学化学与分子工程学院结构可控先进功能材料及其制备教育部重点实验室

出　　处：《光散射学报》2019年第4期367-372,共6页The Journal of Light Scattering

基　　金：国家自然科学基金项目(No.21777041,21575041)

摘　　要：碱性磷酸酶(ALP)是一种重要的酶类生物标志物,其非正常表达与肝功能紊乱、骨病等疾病联系紧密。本研究建立了一种基于表面增强拉曼光谱(SERS)的ALP活性检测新方法,该方法合成了探针分子B-MDP,并利用探针分子磷酸根可被ALP催化水解为酚羟基从而引起SERS谱图明显变化的机制,完成ALP活性的分析检测。得益于SERS技术能够提供分子指纹谱图以及催化反应特异性的复合优点,B-MDP对ALP具有高选择性;同时,该方法可在ALP活性为0.01~0.5 mU/mL和0.5~10 mU/mL范围内呈现良好的线性关系,检出限为30x0E䥺SymbolmA@0x0FU/mL。此外,该方法对血清样品中ALP的加标回收率为98.98%~109.74%,利用该方法也可进行Na 3VO 4对ALP活性的抑制能力评估。结果表明,该方法选择性好、灵敏度高、线性范围宽,可为ALP相关疾病的诊断及其抑制剂筛选提供新的分析方法参考。Alkaline phosphatase(ALP)is a critical enzyme biomarker in living organisms,its abnormal expression has close relationship with liver dysfunction,bone disease and so on.In this work,a novel method based on surface enhanced Raman spectroscopy(SERS)is proposed to detect ALP activity by developing a probe bis-s-s’-(3-mercaptophenyl disodium phosphate)(B-MDP).The detection can be realized by the significant differences in SERS spectra between the probe and its hydrolysis product which is triggered by ALP.Owing to the comprehensive superiority integrating the specificity of catalytical action with the fingerprinting feature of SERS technique,the SERS probe shows high selectivity towards ALP.In addition,the proposed approach demonstrates good linear relationships in the range of 0.01-0.5 mU/mL and 0.5-10 mU/mL with low limit of detection at 30x0E䥺SymbolmA@0x0FU/mL(S/N=3).Furthermore,the proposed SERS method performs well on detecting ALP in serum and evaluating ALP inhibitor,thereby showing a great promising for the early diagnosis and drug screening of ALP-associated diseases.

关 键 词：表面增强拉曼光谱 碱性磷酸酶 探针分子 活性检测 血清 

分 类 号：O43[机械工程—光学工程]