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| 制备蛋白A单分子膜层装配抗体构建肿瘤标志物检测芯片 | |
| 引用本文： | 黄承洪,王春霞,朱伟,侯长军,霍丹群,陈艳艳,靳刚.制备蛋白A单分子膜层装配抗体构建肿瘤标志物检测芯片[J].科学通报,2013,58(7):575-579. |
| 作者姓名： | 黄承洪 王春霞 朱伟 侯长军 霍丹群 陈艳艳 靳刚 |
| 作者单位： | 重庆大学教育部生物流变学重点实验室, 重庆 400044; 中国科学院苏州纳米技术与纳米仿生研究所, 苏州 215123; 山东省医学科学院放射医学研究所, 济南 250062; 中国科学院力学研究所, 北京 100190 |
| 基金项目： | 国家科技支撑计划(2012BAI19B03)、国家自然科学基金(81171414,31171684,11274345)、中国博士后科学基金(2011M500143)、高等学校博士学科点专项科研基金(20090191110030)和重庆大学大型仪器设备开放基金资助 |
| 摘 要： | 免疫分析方法主要依赖于固定在固相基底上的抗体对相应抗原的特异性识别, 抗体在固相基底上固定后最大程度保持生物学活性是设计免疫分析方法的关键技术. 本研究利用在疏水性硅基底表面制备蛋白A单分子膜层可以特异结合抗体的Fc端, 进而实现对抗体的定向化装配, 进一步构建格式化阵列用于肿瘤标志物检测. 研究结果表明, 制备的蛋白A单分子膜层的厚度为1.8±0.6 nm, 抗体可以经蛋白A单分子膜层实现定向化装配, 由此设计的传感阵列检测肿瘤标志物甲胎蛋白(alpha-fetoprotein, AFP)可以实现1.0 ng/mL的灵敏度, 血清检测结果与电化学法(ECLIA)测定进行统计学检验没有显著性差异(P>0.05). |
| 关 键 词： | 蛋白A 单分子层 成像椭偏 甲胎蛋白 |
| 收稿时间： | 2012-05-07 |
Preparation of single molecular film of protein A to assemble antibody for construction of biomarker chip |
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| HUANG ChengHong,WANG ChunXia,ZHU Wei,HOU ChangJun,HUO DanQun,CHEN YanYan,JIN Gang.Preparation of single molecular film of protein A to assemble antibody for construction of biomarker chip[J].Chinese Science Bulletin,2013,58(7):575-579. | |
| Authors: | HUANG ChengHong WANG ChunXia ZHU Wei HOU ChangJun HUO DanQun CHEN YanYan JIN Gang |
| Institution: | 1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, Bioengineering College, Chongqing University, Chongqing 400044, China; 2. Suzhou Institute of Nano-tech and Nano-bionics, Chinese Academy of Sciences, Suzhou 215123, China; 3. The Radiation Medical Institute, Shandong Academy of Medical Sciences, Jinan 250062, China; 4. Institute of Mechanics, Chinese Academy of Sciences, Beijing 100190, China |
| Abstract: | Immunoassay is mainly performed by the immobilized antibody on solid substrate to specifically recognize the corresponding antigen. The key technique is the immobilization of antibody on solid substrate to preserve its biological activities. Here we utilize the protein A to prepare single molecular film on hydrophobic silicon substrate, which can realize site-directed immobilization of antibody by the combination of its Fc fragment for biomarker detection through constructing array format. The results showed that the thickness of single molecular film of protein A measured by imaging ellipsometry is approximately 1.8±0.6 nm and it could be used for site-directed immobilization of antibody. The designed sensing array for AFP detection possesses high sensitivity of 1.0 ng/mL. No statistical difference is found (P>0.05) compared with that of ECLIA. |
| Keywords: | protein A single molecular film imaging ellipsometry &alpha -fetoprotein |
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