期刊
BIOSENSORS & BIOELECTRONICS
卷 54, 期 -, 页码 199-206出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2013.10.044
关键词
Sandwich electrochemiluminescence immunosensor; Surface imprinting; Epitope imprinting; Human immunodeficiency virus type 1 antibody; Horseradish peroxidase; Luminol
类别
资金
- National Natural Science Foundation of China [30901367]
- Natural Science Foundation of Zhejiang [LY13C200017, LY12C20004, LY12B01005]
- Science and Technology Project of Zhejiang [2012C23101, 2011C23126]
- Natural Science Foundation of Ningbo City [2011B82014, 2013A610241]
- K.C. Wong Magna Fund in Ningbo University
In this report, a rapid and cost-effective sandwich electrochemiluminescence (ECL) immiinosensor was constructed for the ultrasensitive detection of human immunodeficiency virus type 1 antibody (anti-HIV-1) using magnetic molecularly imprinted polymers (MMIPs) as capture probes by combining surface and epitope imprinting techniques and antigen conjugated with horseradish peroxidase (HRP-HIV-1) as labels. First, 3-aminobenzeneboronic acid (APBA) was used as the functional monomer and cross-linking reagent, which was polymerized on the surface of silicate-coated magnetic iron oxide nanoparticles (Fe3O4@SiO2 NPs) in the presence of human immunoglobulin G (HIgG), as the template exhibiting the same Pc region but different Fab region to anti-HIV-1 after the addition of the initiator, ammonium persulfate. This process resulted in grafting a hydrophilic molecularly imprinted polymer (MIP) film on the Fe3O4@SiO2 NPs. Thus, MMIPs, which could be reused after eluting the template, were used to recognize and enrich ultra-trace levels of anti-HIV-1. Subsequently, a novel sandwich ECL immunosensor was formed through the immunoreaction between MMIPs conjugated with varied concentrations of anti-HIV-1 and HRP-HIV-1. By the catalysis of HRP immobilized onto HRP-HIV-1 on the ECL system of Luminol-H2O2, a linear response range of the anti-HIV-1 dilution ratio (standard positive serum) was achieved from 1:20,000 to 1:50, with a detection limit of 1:60,000 (S/N=3). The developed method provides a low-cost, simple, and sensitive way for the early diagnosis of HIV infected patients. (C) 2013 Elsevier B.V. All rights reserved.
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