多肽自组装膜用于凝血酶的检测  

Application of Peptide Self-assembled Multilayers on Detection of Thrombin

作　　者：李淑淳 李芳[1] 刘斌[1] 孙向英[1] 

机构地区：[1]华侨大学材料科学与工程学院,厦门361021

出　　处：《分析化学》2014年第7期1044-1048,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.21275059;21175049);福建省自然科学基金(Nos.2012J01044;2011J01049);国务院侨务科研基金(No.11QZR12);中央高校基本科研业务费专项资金(Nos.JB-2R1118;JB-ZR1215)资助~~

摘　　要：利用自组装膜技术在石英片表面构建了多肽自组装膜。考察了组装液浓度和组装时间对多肽自组装膜组装效果的影响,并用紫外可见吸收光谱仪和扫描电镜对该膜进行了表征。研究结果表明,当γ-氨基丙基三乙氧基硅烷水溶液浓度为1%（V/V）,组装时间为3 h；金纳米浓度为2.4×10-4 mol/L,组装时间为12 h；多肽浓度为1×10-4 mol/L,组装时间为12 h时,组装效果最好。基于凝血酶可特异性酶切多肽中精氨酸和甘氨酸之间的肽键,建立了一种测定微量凝血酶的新方法,方法的线性响应范围为2.8×10-12~9.9×10-10 mol/L,检出限为1.4×10-12 mol/L。此膜用于血清样品中凝血酶的测定,回收率为91.6%~107.6%。Peptide self-assembled multilayers （ SAMs ） was coated onto the quartz surface. The assembly conditions, such as the assembly agent concentration and assembly time, were examined. The SAMs was characterized via UV-vis absorption spectrometry and scanning electron microscope. Our results showed that the optimal concentration and assembly time for the 3-aminopropyl-triethoxysilane aqueous were 1% （ V/V ） and 3 h respectively, and those of gold nanoparticles were 2. 4 × 10-4 mol/L and 12 h, respectively, while those of peptide solution were 1 × 10-4 mol/L and 12 h, respectively. A new sensitive method, based on the theory that peptide could be cleaved at the site of Arg-Gly by thrombin, was established to detect thrombin. In addition, a good linear relationship was obtained in the range from 2. 8×10-12 mol/L to 9. 9×10-10 mol/L, and the detection limit was 1. 4×10-12 mol/L. The peptide self-assembled multilayers were also used in the analysis of blood serum samples, and the recovery rate was within the range from 91. 6% to 107. 6%.

关 键 词：多肽 自组装膜 凝血酶 荧光 金纳米 

分 类 号：R446.11[医药卫生—诊断学] O657[医药卫生—临床医学]