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基于温度可控微系统的珠亲和层析法用于生物标志物检测。

Bead affinity chromatography in a temperature-controllable microsystem for biomarker detection.

机构信息

School of Electrical Engineering and Computer Science, Seoul National University, KwanAk Ku, Seoul, South Korea.

出版信息

Anal Bioanal Chem. 2012 Nov;404(8):2267-75. doi: 10.1007/s00216-012-6380-1. Epub 2012 Sep 18.

Abstract

This paper describes a temperature-controllable bead affinity chromatography (BAC) in a microsystem for biomarker detection, and preparing samples for matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) analysis. Cancer marker proteins were captured in the microsystem by BAC with RNA aptamer-immobilized microbeads. The captured proteins were then denatured and released from the microbeads by controlling temperature. The microsystem consists of a microreactor for trapping microbeads and a temperature control unit for thermal treatment of the trapped beads. We used polymethylsilxoane or single crystalline silicon in fabricating two different types of reaction chamber to compare the differences in performance originated from the materials. Carcinoembryonic antigen was concentrated and purified from human serum using the microsystem and detected by MALDI-TOF MS to demonstrate the usefulness of the microsystem. The microsystem simplifies a sample preparation process required for protein analysis and cancer biomarker detection, which will accelerate the process of cancer research.

摘要

本文描述了一种用于生物标志物检测的微系统中温度可控的珠状亲和层析(BAC),并为基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)分析制备样品。通过将 RNA 适体固定在微珠上的 BAC,在微系统中捕获癌症标志物蛋白。然后通过控制温度使捕获的蛋白质变性并从微珠上释放出来。微系统由一个用于捕获微珠的微反应器和一个用于对捕获的微珠进行热处理的温度控制单元组成。我们使用聚甲基硅氧烷或单晶硅制造了两种不同类型的反应室,以比较源自材料的性能差异。使用微系统从人血清中浓缩和纯化癌胚抗原,并通过 MALDI-TOF MS 进行检测,以证明微系统的有用性。该微系统简化了蛋白质分析和癌症生物标志物检测所需的样品制备过程,这将加速癌症研究的进程。

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