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基于芯片的免疫亲和毛细管电泳法分析新生儿干血斑样本中的炎症介质

Analysis of Inflammatory Mediators in Newborn Dried Blood Spot Samples by Chip-Based Immunoaffinity Capillary Electrophoresis.

作者信息

Phillips Terry M, Wellner Edward F

机构信息

Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, VA, USA.

Department of Pharmaceutics, Virginia Commonwealth University, Washington, DC, USA.

出版信息

Methods Mol Biol. 2019;1972:185-198. doi: 10.1007/978-1-4939-9213-3_13.

Abstract

A chip-based immunoaffinity capillary electrophoresis (ICE) system has been developed for measuring inflammatory mediators in dried blood samples routinely taken from newborn babies. A defined area of each dried blood spot was removed from the sample card and its contents eluted. The recovered eluates were injected into the chip and the analytes of interest isolated by the immunoaffinity disk within the chip. The captured analytes were labeled in-situ with a red light-emitting laser dye and electro-eluted into the chip separation channel. Electrophoretic separation of all of the analytes was achieved within 2.0 min with quantification of each peak being performed by online LIF detection and integration of each peak area. The degree of accuracy and precision achieved by the chip-based system is comparable to conventional immunoassays and the system is robust enough to be applied to the analysis of clinical samples.

摘要

一种基于芯片的免疫亲和毛细管电泳(ICE)系统已被开发出来,用于测量从新生儿常规采集的干血样本中的炎症介质。从样本卡上移除每个干血斑的指定区域,并洗脱其内容物。将回收的洗脱液注入芯片,通过芯片内的免疫亲和盘分离出感兴趣的分析物。捕获的分析物用红色发光激光染料原位标记,然后电洗脱到芯片分离通道中。所有分析物在2.0分钟内实现电泳分离,通过在线激光诱导荧光检测和每个峰面积积分对每个峰进行定量。基于芯片的系统所达到的准确度和精密度与传统免疫测定相当,并且该系统足够稳健,可应用于临床样本分析。

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