An enhanced ELISA based on modified colloidal gold nanoparticles for the detection of Pb(II) |
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Authors: | Zhou Yu Tian Xiang-Li Li Yan-Song Pan Feng-Guang Zhang Yuan-Yuan Zhang Jun-Hui Yang Li Wang Xin-Rui Ren Hong-Lin Lu Shi-Ying Li Zhao-Hui Chen Qi-Jun Liu Zeng-Shan Liu Jing-Qiu |
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Affiliation: | Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, Jilin University, Xi-an Road 5333, Changchun, Jilin 130062, PR China. zhouyu69@sina.com |
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Abstract: | A novel probe based on colloidal gold nanoparticles (AuNPs) modified with goat anti-mouse IgG and horseradish peroxidase (HRP) was synthesized and an enhanced enzyme-linked immunosorbent assay (ELISA) based on the probe was developed. In the assay, the synthesized probe is bound with a monoclonal antibody (McAb) which is competitively bound by coated BSA-ITCBE-Pb(II) on plate and Pb(II) in samples. The HRP, used here for signal amplification catalytically oxidize the substrate and generate optical signals that is related to the concentration of Pb(II) and can be measured spectrophotometrically. For the monodisperse AuNPs having high surface areas, it can be conjugated with more amount of HRP than that of IgG. Therefore, compared with traditional ELISA, the signal amplification of catalytically oxidized substrate was enhanced. The detection limit for this novel modified AuNPs probe-based assay was 9 pg mL(-1). The recoveries obtained by standard Pb(II) addition to real samples, including a commercial mineral water, tap water, and lake water were all from 94.9% to 102.9%. And the coefficient of variation (CV) value of all samples was less than 10%. The results indicated that the enhanced assay gave higher sensitivity and reliable reproducibility. It could provide a general detection format for low-molecular weight contaminants. |
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