量子点标记的斑点免疫渗滤分析定量检测cTnI

高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (11): 2180.

量子点标记的斑点免疫渗滤分析定量检测cTnI

范佳^1,3, 宋健², 毕丽荣⁵, 周广宇⁵, 张皓⁴, 魏景艳^1,3, 杨柏⁴

1. 吉林大学药学院,
2. 电子科学与工程学院,
3. 分子酶学工程教育部重点实验室, 长春 130021;
4. 吉林大学化学学院, 长春 130012;
5. 吉林大学中日联谊医院, 长春 130033

收稿日期:2008-11-24 出版日期:2009-11-10 发布日期:2009-11-10
通讯作者: 魏景艳, 女, 博士, 教授, 博士生导师, 主要从事生物化学与分子生物学研究. E-mail: jingyanwei@yahoo.com.cn
基金资助:
吉林省科学技术厅基金(批准号: 200705380)、中国博士后基金(批准号: 2004036395)和吉林大学本科生创新性实验计划(批准号: 2007C3268)资助.

Dot Immunofiltration Assay for Quantitative Detecting cTnI Using Quantum Dots

FAN Jia^1,3, SONG Jian², BI Li-Rong⁵, ZHOU Guang-Yu⁵, ZHANG Hao⁴, WEI Jing-Yan^1,3*, YANG Bai⁴

1. College of Pharmacy,
2. College of Electronic Science and Engineering,
3. Key Laboratory of Molecular Enzymology and Engineering, Ministry of Education, Jilin University, Changchun 130021, China;
4. College of Chemistry, Jilin University, Changchun 130012, China;
5. China-Japan Union Hsopital, Jilin University, Changchun 130033, China

Received:2008-11-24 Online:2009-11-10 Published:2009-11-10
Contact: WEI Jing-Yan. E-mail: jingyanwei@yahoo.com.cn
Supported by:
吉林省科学技术厅基金(批准号: 200705380)、中国博士后基金(批准号: 2004036395)和吉林大学本科生创新性实验计划(批准号: 2007C3268)资助.

摘要/Abstract

摘要：

利用量子点良好的光谱特征和光化学稳定性, 结合免疫分析技术, 对心肌肌钙蛋白I(cTnI)特异性进行定量检测. 用量子点标记cTnI的单克隆抗体(2F11), 通过SDS-PAGE电泳证明标记成功. 斑点免疫膜渗滤法证明标记后的2F11仍具有良好的生物学活性, 再将标记并纯化后的2F11与NC膜上不同浓度的cTnI进行免疫反应, 使用ImageMaster图像分析软件对膜上荧光斑点图像进行定量分析. 应用此方法测得cTnI的浓度和斑点处相对荧光值有良好的线性关系(R²=0.9966), 最低检出值为120 ng.

关键词: 心肌肌钙蛋白I; 特异性抗心肌肌钙蛋白I单克隆抗体; 斑点免疫渗滤; 量子点

Abstract:

Quantum dots have physical and optical properties that make them useful tools for high-resolution labeling immunoassay. In this work, a rapid and simple method of quantitative immunoassay for Cardiac troponin I(cTnI) was developed with quantum dots-labeled antibodies. The monoclonal antibodies of cTnI(2F11) could be labeled with CdTe quantum dots and the coupled product(CdTe-2F11) were characterized by SDS-PAGE. The result of immunofiltration assay indicats that the CdTe-2F11 maintains the antibody activity. The cTnI at the different concentrations in NC membrane could react with CdTe-2F11 and be detected with ImageMaster to analyze the fluorescence intensity of the immunodotting. The results show that the detection limit of cTnI is 120 ng, and there is a good linear relation between concentration of cTnI and the fluorescence intensity(R²=0.9966).

Key words: Cardiac troponin I; Monoclonal antibody to cardiac troponin I; Dot immunofiltration assay; Quantum dot

TrendMD:

范佳, 宋健, 毕丽荣, 周广宇, 张皓, 魏景艳, 杨柏. 量子点标记的斑点免疫渗滤分析定量检测cTnI. 高等学校化学学报, 2009, 30(11): 2180.

FAN , SONG Jian, BI Li-Rong, ZHOU Guang-Yu, ZHANG Hao, WEI Jing-Yan. Dot Immunofiltration Assay for Quantitative Detecting cTnI Using Quantum Dots. Chem. J. Chinese Universities, 2009, 30(11): 2180.

参考文献

[1]Bruchez M. J., Moronne M., Alivisatos A. P., et al.. Science[J], 1998, 281(5385): 2013—2016
[2]Chan W., Nie S. M.. Science[J], 1998, 281(5385): 2016—2018
[3]Seydel C.. Science[J], 2003, 300(5626): 1659
[4]Sun B. Q., Xie W. Z., Yi G. S., et al.. J. Immunol. Methods[J], 2001, 249(1/2): 85—89
[5]Goldman E. R., Anderson G. P., Tran P. T., et al.. Anal. Chem.[J], 2002, 74(4): 841—847
[6]Wang S. P., Mamedova N., Kotov N. A., et al.. Nano Lett.[J], 2002, 2(8): 817—822
[7]Lingerfelt B. M., Mattoussi H., Goldman E. R., et al.. Anal. Chem.[J], 2003, 75(16): 4043—4049
[8]Wu X. Y., Liu H. J., Liu J. Q., et al.. Nat. Biotechnol.[J], 2003, 21(1): 41—46
[9]Alivisatos A. P., Gu W., Larabell C.. Annu. Rev. Biomed. Eng.[J], 2005, 7: 55—76
[10]Ghazani A. A., Lee J. A., Klostranec J., et al.. Nano Lett.[J], 2006, 6(12): 2881—2886
[11]Barton P., Cullen M. E., Townsend P. J., et al.. Genomics[J], 1999, 57(1): 102—109
[12]Zhang H., Yang B.. Thin Solid Films[J], 2002, 418(2): 169—174
[13]LIN Zhang-Bi(林章碧), SU Xing-Guang(苏星光), ZHANG Hao(张皓), et al.. Chem. J. Chinese Universities(高等学校化学学报)[J], 2003, 24(2): 216—220
[14]WEI Jing-Yan(魏景艳), QIAN Li-Na(钱丽娜), LU Jie(芦杰), et al.. Chin. J. Biologicals(中国生物制品学杂志)[J], 2002, 15(5): 29—31
[15]WEI Jing-Yan(魏景艳), FU Ping-Ping(付平平), SONG Ang(宋昂). Chin. J. Biologicals(中国生物制品学杂志)[J], 2000, 13(4): 218—220
[16]Wei J. Y., Song D. Q., Bu L. S., et al.. Chem. Res. Chinese Universitis[J], 2003, 19(2): 183—189
[17]WEI Jing-Yan(魏景艳), BU Li-Sha(卜丽莎), DONG Yu-Jun(董玉军), et al.. Chin. J. Laboratory Medicine(中华检验医学杂志)[J], 2003, 26(5): 1—3
[18]SONG Jian(宋健), FAN Jia(范佳), SONG Da-Qian(宋大千), et al.. Chem. J. Chinese Universities(高等学校化学学报)[J], 2009, 30(10): 1940—1944
[19]Hua X. F., Liu T. C., Cao Y. C., et al.. Anal. Bioanal. Chem.[J], 2006, 386(6): 1665—1671