On-line electrophoretic sample clean-up for sensitive and reproducible μCE immunoassay.
On-line electrophoretic sample clean-up for sensitive and reproducible μCE immunoassay.
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在线电泳样品净化，用于灵敏且可重复的 μCE 免疫分析。
DOI：
10.1039/c2lc21111d
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发表时间：
2012
期刊：
影响因子：
6.1
通讯作者：
Luke P. Lee
中科院分区：
文献类型：
--
作者：
Qiong Pan;SoonGweon Hong;Xiaocui Zhu;Meiping Zhao;Luke P. Lee
我们报道了一种新的在线电泳样品净化方法，用于人血清中低丰度蛋白质的高灵敏度和可重复性的微芯片电泳（μCE）免疫分析。该方法利用场放大样品堆积对具有不同电泳迁移率的分子的差异效应。通过简单的电压控制从上样通道中去除大的干扰蛋白，从而选择性浓缩并将较小的目标分析物注入分离通道。作为概念验证，开发了一种无抗体进样模式，用于血清样本中人胰岛素样生长因子-I（IGF-I）的直接μCE免疫测定，无需任何额外的纯化步骤。IGF-I峰清晰、尖锐，背景低，重现性好。此外，通过在进行μCE检测前向样品缓冲液中加入浓度为50%的乙醇，进一步提高了测定灵敏度。IGF-I的检测下限达到0.68 ng mL（-1），总信号增强因子为2750。所建立的在线电泳样品净化方法可能会在其他基于微芯片的临床和生物学用途的高通量分析平台的开发中找到广泛的应用。
We report a novel on-line electrophoretic sample clean-up approach for highly sensitive and reproducible microchip electrophoretic (μCE) immunoassay of low-abundance proteins in human serum. The method takes advantage of the differential effect of field-amplified sample stacking on molecules with different electrophoretic mobility. Large interfering proteins are removed from the loading channel by simple voltage control, resulting in selective concentration and injection of smaller target analytes to the separation channel. As a proof of concept, an antibody-free injection mode was developed for direct μCE immunoassay of human insulin-like growth factor-I (IGF-I) in serum samples without any additional purification steps. Clear and sharp peaks were obtained for IGF-I with low background and excellent reproducibility. Besides, the assay sensitivity was further increased by addition of ethanol to the sample buffer at a concentration of 50% right before performing the μCE detection. The lower limit of detection of IGF-I achieved 0.68 ng mL(-1), with an overall signal enhancement factor of 2750. The established on-line electrophoretic sample clean-up approach may find wide applications in the development of other microchip-based high-throughput analytical platforms for clinical and biological use.
影响因子：
7.4
作者：
SCHULTZ, NM;HUANG, L;KENNEDY, RT
通讯作者：
KENNEDY, RT
影响因子：
7.4
作者：
Pan, Qiong;Zhao, Meiping;Liu, Shaorong
通讯作者：
Liu, Shaorong