Single-drop analysis of various proteases in a cancer cell lysate using a capillary-assembled microchip
Single-drop analysis of various proteases in a cancer cell lysate using a capillary-assembled microchip
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DOI：
10.1007/s00216-008-2105-x
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发表时间：
2008-08-01
影响因子：
4.3
通讯作者：
Hisamoto, Hideaki
中科院分区：
文献类型：
--
作者：
Henares, Terence G.;Mizutani, Fumio;Hisamoto, Hideaki
提出了对两种不同的真实样品溶液（2μL）进行单滴分析，同时监测单个微流体装置上两组十种不同蛋白酶的活性。该装置被称为毛细管组装微芯片（CAs-CHIP），是通过在聚二甲基硅氧烷（PDMS）晶格微通道中嵌入方形玻璃传感毛细管（试剂释放毛细管，RRC）来制造的，并用于此目的。首先，通过测量单个 CAs-CHIP 上 20 个 caspase-3 传感毛细管的荧光响应来评估性能可靠性，获得 1.5-8.2（% RSD，n=5 或 10）的相对标准偏差。这表明通过使用嵌入多个 RRC 的单个 CAs-CHIP 可以实现精确的多重蛋白酶活性传感。然后，使用单个 CAs-CHIP，同时对经细胞死亡诱导药物阿霉素处理和未处理的宫颈癌 (HeLa) 细胞裂解液中 10 种不同的半胱天冬酶/蛋白酶的活性进行真实样品分析，并观察到这两个样品之间酶活性的显着差异。这些结果表明 CAs-CHIP 在药物发现领域的有用性。
Single-drop analysis of two different real sample solutions (2 mu L) while simultaneously monitoring the activity of two sets of ten different proteases on a single microfluidic device is presented. The device, called a capillary-assembled microchip (CAs-CHIP), is fabricated by embedding square glass sensing capillaries (reagent-release capillaries, RRC) in the polydimethylsiloxane (PDMS) lattice microchannel, and used for that purpose. First, the performance reliability was evaluated by measuring the fluorescence response of twenty caspase-3-sensing capillaries on a single CAs-CHIP, and a relative standard deviation of 1.5-8.2 (% RSD, n=5 or 10) was obtained. This suggests that precise multiplexed protease-activity sensing is possible by using a single CAs-CHIP with multiple RRCs embedded. Then, using a single CAs-CHIP, real sample analysis of the activity of ten different caspases/proteases in cervical cancer (HeLa) cell lysate treated and untreated with the cell-death-inducer drug, doxorubicin, was simultaneously carried out, and a significant difference in enzyme activity between these two samples was observed. These results suggested the usefulness of the CAs-CHIP in the field of drug discovery.