Enhanced colorimetric immunoassay accompanying with enzyme cascade amplification strategy for ultrasensitive detection of low-abundance protein.

Enhanced colorimetric immunoassay accompanying with enzyme cascade amplification strategy for ultrasensitive detection of low-abundance protein.
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增强的比色免疫测定结合酶级联放大策略,用于低丰度蛋白质的超灵敏检测。

DOI:
10.1038/srep03966
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发表时间:
2014-02-10
期刊:
影响因子:
4.6
通讯作者:
Tang D
Tang D
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Gao Z;Hou L;Xu M;Tang D

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基于酶标记的方法已被开发用于比色免疫分析,但大多数方法灵敏度较低,不适合常规使用。在这里,我们设计了一种结合酶-级联扩增策略(ECAS-CIA)的前列腺特异性抗原(PSA)的增强比色免疫分析方法。在靶标PSA存在下,标记在二抗上的碱性磷酸酶催化钯纳米结构的形成,该纳米结构催化3,3‘,5,5’-四甲基联苯胺-H_2O_2体系生成有色产物,从而导致信号级联放大。结果表明,ECAS-CIA对PSA有良好的响应,可检测浓度低至0.05 ng mL−-1的PSA,批内和批间变异系数分别小于9.5%和10.7%。此外,该方法被用于临床血清标本的分析,与PSA酶联免疫试剂盒获得的结果一致。重要的是,ECAS-CIA为蛋白质诊断和生物安全开辟了一个新的领域。
Methods based on enzyme labels have been developed for colorimetric immunoassays, but most involve poor sensitivity and are unsuitable for routine use. Herein, we design an enhanced colorimetric immunoassay for prostate-specific antigen (PSA) coupling with an enzyme-cascade-amplification strategy (ECAS-CIA). In the presence of target PSA, the labeled alkaline phosphatase on secondary antibody catalyzes the formation of palladium nanostructures, which catalyze 3,3′,5,5′-tetramethylbenzidine-H2O2 system to produce the colored products, thus resulting in the signal cascade amplification. Results indicated that the ECAS-CIA presents good responses toward PSA, and allows detection of PSA at a concentration as low as 0.05 ng mL−1. Intra- and inter-assay coefficients of variation are below 9.5% and 10.7%, respectively. Additionally, the methodology is validated for analysis of clinical serum specimens with consistent results obtained by PSA ELISA kit. Importantly, the ECAS-CIA opens a new horizon for protein diagnostics and biosecurity.
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