Development of a micro total analysis system incorporating chemiluminescence detection and application to detection of cancer markers
Development of a micro total analysis system incorporating chemiluminescence detection and application to detection of cancer markers
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DOI：
10.1021/ac040133t
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发表时间：
2005-03-15
影响因子：
7.4
通讯作者：
Nakajima, R
中科院分区：
文献类型：
--
作者：
Tsukagoshi, K;Jinno, N;Nakajima, R
采用异硫氰鲁米诺（ILITC）作为化学发光标记试剂，微过氧化物酶作为催化剂，过氧化氢作为氧化剂的化学发光反应，建立了微全分析系统（mu-TAS）。该分析系统在微芯片上进行以下三个过程：高选择性的免疫反应，样品塞的形成和运输的电泳，和高灵敏度的化学发光检测。将这三个过程复杂地集成到微芯片上以得到mu-TAS。该微芯片包含两个在交叉点交叉的微通道，而微通道的末端进入四个水库。作为第一步，使用抗体固定化玻璃珠进行免疫反应。将玻璃珠与抗原（分析物）和已知量的ILITC标记抗原一起沿着放置在一个储器中，以建立竞争性免疫反应。对于电泳，作为第二个过程，免疫反应后的反应物被电泳地进料到交叉点中，导致样品塞。然后将样品塞移动到另一个含有过氧化氢溶液的储存器中。此时，进行化学发光检测作为第三过程：标记抗原与过氧化氢和迁移缓冲液中包含的催化剂混合以产生化学发光。化学发光由位于储液器下方的光电倍增管检测。本文描述的mu-TAS能够以高选择性和灵敏度测定人血清中作为癌症标志物的人血清白蛋白或免疫抑制酸性蛋白。
We developed a micro total analysis system (mu-TAS) incorporating chemiluminescence detection, in which the chemiluminescence reaction of isoluminol isothiocyanato (ILITC) (as a chemiluminescence reagent for labeling)-microperoxidase (as a catalyst)-hydrogen peroxide (as an oxidant) was adopted. The analysis system performed the following three processes on a microchip: immune reaction for high selectivity, electrophoresis for formation and transportation of the sample plug, and chemiluminescence detection for high sensitivity. The three processes were compactly integrated onto the microchip to give the mu-TAS. The microchip contained two microchannels that crossed at an intersection, while the ends of the microchannels accessed four reservoirs. As the first process, the immune reaction was performed using an antibody-immobilized glass bead. The glass bead was placed in one of the reservoirs along with antigen (analyte) and a known amount of ILITC-labeled antigen to set up a competitive immune reaction. For electrophoresis, as the second process, the reactant after the immune reaction was fed electrophoretically into the intersection resulting in a sample plug. The sample plug was then moved into another reservoir containing hydrogen peroxide solution. At this point, chemiluminescence detection was performed as the third process: the labeled antigen mixed with the hydrogen peroxide and the catalyst included in the migration buffer to produce chemiluminescence. Chemiluminescence was detected by a photomultiplier tube located under the reservoir. The mu-TAS described here was capable of determining, with high selectivity and sensitivity, human serum albumin or immunosuppressive acidic protein as a cancer marker in human serum.