Fast and sensitive detection of protein concentration in mild environments
Fast and sensitive detection of protein concentration in mild environments
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在温和环境下快速灵敏地检测蛋白质浓度
DOI：
10.1016/j.talanta.2014.12.046
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发表时间：
2015-04-01
期刊：
影响因子：
6.1
通讯作者：
Xia, Xing-Hua
中科院分区：
文献类型：
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作者：
Wang, Chen;Zhou, Yue;Xia, Xing-Hua
温和环境中蛋白质浓度的测定在临床诊断和生物检测中具有重要意义。建立了一种简便、快速、灵敏的中性溶液（pH7.0）中蛋白质定量测定方法。该测定基于样品蛋白和荧光标记的狗血清白蛋白（FITC-DSA）在金纳米颗粒（AuNP）上的竞争吸附。当竞争性FITC-DSA分子加入到样品蛋白缀合的AuNPs的混合溶液中时，它们将竞争AuNPs的活性位点，导致由于AuNPs通过荧光共振能量转移（FRET）的猝灭效应而导致荧光强度降低。因此，可以实现样品蛋白质浓度的定量测定。在最佳条件下，溶液的荧光强度的降低与样品蛋白质的浓度有关，并且在5 min内可以达到0.01 μ g/mL BSA的低检测限。为了验证我们的策略在实际应用中的有效性，使用所提出的方法测定了人血清中的总蛋白质含量。结果表明，该方法简便、准确、灵敏度高，适用于生物测定中蛋白质的检测。(C)2015 Elsevier B. V.版权所有。
Determination of protein concentration in mild environments is of great significance in the clinic diagnose and bioassay. Herein, a simple, fast and sensitive method for protein quantitative determination in neutral solution (pH 7.0) is developed. This assay is based on competition adsorption of the sample protein and fluorescently labeled dog serum albumin (FITC-DSA) onto gold nanoparticles (AuNPs). As the competitor FITC-DSA molecules are added into the mixture solution of sample protein conjugated AuNPs, they will compete for active sites of AuNPs, resulting in decrease in fluorescence intensity due to the quenching effect of AuNPs via fluorescence resonance energy transfer (FRET). Thus, quantitative determination of sample protein concentration can be achieved. Under the optimum conditions, the decrease in fluorescence intensity of the solution is related to the concentration of sample protein and a low detection limit of 0.01 mu g/mL BSA can be achieved in 5 min. For the validation of our strategy in practical applications, the total protein content in human serum was determined using the as-proposed method. The result is in well agreement with that of measured by other conventional methods, suggesting a simple, accurate, and mild approach for protein detection in bioassay. (C) 2015 Elsevier B.V. All rights reserved.