Aldo Paulomi, Marusov Gregory, Svancara Danielle, David James, Mor Gil
Division of Reproductive Sciences, Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine. New Haven, CT, USA.
ProteinSimple, San Jose, CA, USA.
Am J Reprod Immunol. 2016 Jun;75(6):678-93. doi: 10.1111/aji.12512. Epub 2016 May 11.
Quantitative measurement of proteins in bodily fluids or cellular preparations is critical for the evaluation of biomarkers or the study of complex cellular processes. While immunoassays are the most common quantitative approach used so far, they are not practical for the evaluation of multiple proteins. Microfluidic technology allows a fine spatial control in immobilizing proteins and biomolecules inside microchannels, eliminating cross-reactivity between competing analytes, and allowing rapid and sensitive detection of targeted antigens for multiple applications. We report the characterization and validation of the Simple Plex(™) platform for the detection and quantification of cytokines and chemokines from human and mouse samples.
Cytokine and chemokine expression levels were determined using Simple Plex cartridges from ProteinSimple. Serum samples were obtained from the Yale Biorepository.
Our data demonstrate an excellent correlation between the results obtained with Simple Plex and conventional immunoassays such as ELISA and Luminex.
We describe the characterization and validation of Simple Plex, a novel multi-analyte, automated microfluidic platform that allows the evaluation of cytokines and chemokines from human and mice biological samples. Simple Plex showed significant advantages over traditional approaches in terms of low sample volume requirements, sensitivity and dynamic range, coefficient of variation, and reproducibility.
对体液或细胞制剂中的蛋白质进行定量测量对于生物标志物评估或复杂细胞过程研究至关重要。虽然免疫测定法是迄今为止最常用的定量方法,但它们对于多种蛋白质的评估并不实用。微流控技术能够在微通道内固定蛋白质和生物分子时实现精细的空间控制,消除竞争性分析物之间的交叉反应,并允许对多种应用中的靶向抗原进行快速灵敏的检测。我们报告了用于检测和定量来自人和小鼠样本的细胞因子和趋化因子的Simple Plex™平台的特性及验证情况。
使用ProteinSimple公司的Simple Plex试剂盒测定细胞因子和趋化因子的表达水平。血清样本取自耶鲁生物样本库。
我们的数据表明,Simple Plex获得的结果与酶联免疫吸附测定(ELISA)和Luminex等传统免疫测定法的结果具有良好的相关性。
我们描述了Simple Plex的特性及验证情况,这是一种新型的多分析物自动化微流控平台,可用于评估来自人和小鼠生物样本的细胞因子和趋化因子。在低样本量要求、灵敏度和动态范围、变异系数及重现性方面,Simple Plex相较于传统方法具有显著优势。