Citation: DING Feng-Li, CHENG Qiong, LI Jun-Feng, JIANG Fei-En. Magnetic Nanospheres-based Electrochemical Immunoassay Amplified by Hyperbranched Polymer[J]. Chinese Journal of Analytical Chemistry, ;2012, 40(10): 1514-1518. doi: 10.3724/SP.J.1096.2012.11447 shu

Magnetic Nanospheres-based Electrochemical Immunoassay Amplified by Hyperbranched Polymer

1.
¹ College of Petroleum and Chemical Engineering, Changzhou University, Changzhou 213164, China;

Corresponding author: CHENG Qiong,
Received Date: 23 December 2011
Available Online: 22 April 2012
Fund Project: 本文系浙江省自然科学基金(No.Y405468) (No.Y405468) 浙江省科技计划(No.2007F70008)嘉兴市科技计划项目(No.2008AY2017)资助 (No.2007F70008)嘉兴市科技计划项目(No.2008AY2017)

A novel ultra-sensitive immunoassay for Hepatitis B surface antigen (HBsAg) was proposed. Magnetic nanosphere functionalized with carboxyl group was activated with 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide hydrochloride/N-hydroxy-succinimimide (EDC/NHS), and then Hepatitis B surface antibody (HBsAb) was covalently immobilized on the surface. Hepatitis B secondary antibody labeled with horseradish peroxide (HRP) was covalently linked to the hyperbranched polymer(HBP). The antibody could capture the HBsAg when the antibody-modified magnetic nanosphere was incubated with HBsAg. Then the HBP conjugate was added, and a sandwich immunocomplex formed on the surface of magnetic nanospheres. The nanosphere with sandwich complex was separated magnetically from sample solution, and then incubated in the buffer solution containing 2-amino hydroxy benzene and hydrogen peroxide. The HRP could catalyze the reaction between 2-amino hydroxy benzene and hydrogen peroxide to produce electroactive product 2-hydroxy-3-amino phenoxazine. When using differential pulse voltammetry. the peak current was linear with the concentration of HBsAg in the range of 0.05 to 10 μg/L under the optimum conditions. The detection limit was found to be 0.008 μg/L, and the linear regression equation was I(μA)＝0.140+16.80C(μg/L) with r＝0.9995. This method was applied to analyze real samples.
- Hyperbranched polymer,
- Magnetic nanosphere,
- Hepatitis B surface antigen,
- Immunoassay
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沈阳化工大学材料科学与工程学院沈阳 110142