SERS-based Immunoassay in a Microfluidic System for the Multiplexed Recognition of Interleukins from Blood Plasma: Towards Picogram Detection.

SERS-based Immunoassay in a Microfluidic System for the Multiplexed Recognition of Interleukins from Blood Plasma: Towards Picogram Detection.
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DOI:
10.1038/s41598-017-11152-w
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发表时间:
2017-09-06
期刊:
影响因子:
4.6
通讯作者:
Waluk J
Waluk J
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Kamińska A;Winkler K;Kowalska A;Witkowska E;Szymborski T;Janeczek A;Waluk J

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已经开发了结合到微流体装置中的SERS活性纳米结构,用于快速和多重监测血浆中选定的1型细胞因子(白介素:IL-6、IL-8、IL-18)水平。已经通过使用纳米颗粒进行了多种分析,每个纳米颗粒涂覆有不同的拉曼报告分子:5,5 ′-二硫代-双(2-硝基苯甲酸)(DTNB)、品红(FC)和对巯基苯甲酸(p-MBA)以及特异性抗体。多元统计方法,主成分分析(PCA),应用于分离的三个不同的抗原-抗体复合物编码的三个拉曼报告(FC,p-MBA,和DTNB)在同时多重检测方法。据我们所知,我们还提出了,第一次,一种可能性的多重定量的三种白细胞介素:IL-6,IL-8和IL-18在血浆样品中使用Sers技术。与标准ELISA方法相比,我们的方法提高了检测限。在平行方法中,IL-6、IL-8和IL-18的低检测限估计分别为2.3 pg·ml-1、6.5 pg·ml-1和4.2 pg·ml-1,在同时多重方法中,低检测限估计分别为3.8 pg·ml-1、7.5 pg·ml-1和5.2 pg·ml-1。这证明了分析检查所需的灵敏度和重现性。
SERS-active nanostructures incorporated into a microfluidic device have been developed for rapid and multiplex monitoring of selected Type 1 cytokine (interleukins: IL-6, IL-8, IL-18) levels in blood plasma. Multiple analyses have been performed by using nanoparticles, each coated with different Raman reporter molecules: 5,5′-dithio-bis(2-nitro-benzoic acid) (DTNB), fuchsin (FC), and p-mercatpobenzoic acid (p-MBA) and with specific antibodies. The multivariate statistical method, principal component analysis (PCA), was applied for segregation of three different antigen-antibody complexes encoded by three Raman reporters (FC, p-MBA, and DTNB) during simultaneous multiplexed detection approach. To the best of our knowledge, we have also presented, for the first time, a possibility for multiplexed quantification of three interleukins: IL-6, IL-8, and IL-18 in blood plasma samples using SERS technique. Our method improves the detection limit in comparison to standard ELISA methods. The low detection limits were estimated to be 2.3 pg·ml−1, 6.5 pg·ml−1, and 4.2 pg·ml−1 in a parallel approach, and 3.8 pg·ml−1, 7.5 pg·ml−1, and 5.2 pg·ml−1 in a simultaneous multiplexed method for IL-6, IL-8, and IL-18, respectively. This demonstrated the sensitivity and reproducibility desirable for analytical examinations.
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