高等学校化学学报 ›› 2005, Vol. 26 ›› Issue (3): 441.

• 研究论文 • 上一篇    下一篇

一种基于静电吸附作用的直接检测补体C3新型可再生电容型免疫传感器

吴再生, 李继山, 罗明辉, 沈国励, 俞汝勤   

  1. 湖南大学化学生物传感与计量学国家重点实验室, 化学化工学院 长沙 410082
  • 收稿日期:2004-01-14 出版日期:2005-03-10 发布日期:2005-03-10
  • 通讯作者: 沈国励(1938年出生),男,教授,博士生导师,从事化学与生物传感及其在生化、环境及材料分析中的应用研究.E-mail:glshen@hnu.cn E-mail:glshen@hnu.cn
  • 基金资助:

    国家自然科学基金(批准号:20075006,20375012,20205004);教育部博士点基金(批准号:20010532008)资助.

A Novel Regenerative Capacitive Immunosensor Based on Electrostatic Attraction for Direct Detection of the Complement Ⅲ(C3)

WU Zai-Sheng, LI Ji-Shan, LUO Ming-Hui, SHEN Guo-Li, YU Ru-Qin   

  1. State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China
  • Received:2004-01-14 Online:2005-03-10 Published:2005-03-10

摘要: 通过壳聚糖/褐藻酸钠体系实现抗体在电极上的固定,制得可多次再生的电容型免疫传感器,用于补体C3的检测.先在金电极表面上组装一层半胱胺单分子层,通过戊二醛把壳聚糖修饰在金电极上,再用十二硫醇封闭电极,最后,利用壳聚糖与褐藻酸钠之间的强静电相互作用实现抗体的固定.使用交流阻抗法研究了溶液pH值和离子强度对电极膜层稳定性及对抗体固定性的影响.结果表明,所制备的传感器操作简便,易于再生,电容响应与补体C3浓度在18.2~292.5ng/mL范围内呈线性关系,检出限为9.1ng/mL.

关键词: 静电作用, 电容型免疫传感器, 补体C3

Abstract: A regenerative capacitive immunosensor was proposed for direct detection of the complement Ⅲ(C3) in human serum. Cysteamine selF-assembled monolayers(SAM) firstly were combined on the gold electrode surface, and then a positively-charged chitosan monolayer was attached on the amine-modified SAM via the aid of glutaraldehyde cross-linking. To increase the insulation degree of the layer, the modified electrode was blocked with 1-dodecanethiol ethanol solution. Finally, the natural polysaccharide alginate, a negatively-charge polyelectrolyte, which was previously coupled with the anti-C3 was adsorbed on the chitosan monolayer by strong electrostatic interaction to accomplish the immobilization of the anti-C3. The dependence of chitosan/Au electrode stability on the pH and ion strength was studied with alternating current impedance. The immunosensor fabricated with proposed method can offer high sensitivity and easy regeneration after immunoassay through washing with basic solution without obvious decrease in response. The linear calibration curve is obtained with a detection limit of 9.1 ng/mL and a dynamic response range from 18.2 to 292.5 ng/mL when plotted vs. the logarithm of the antigen concentration. A recovery experiment demonstrates the feasibility of the proposed immunosensor for clinical diagnosis.

Key words: Electrostatic attraction, Capacitive immunosensor, Complement Ⅲ

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