Department of Chemistry, Seoul National University, Seoul 151-747, Republic of Korea.
Talanta. 2013 May 15;109:20-5. doi: 10.1016/j.talanta.2013.03.028. Epub 2013 Mar 20.
An on-chip immunoassay to detect C-reactive protein (CRP) was performed using a polyester-toner (PT) microchip. CRP is a highly conserved plasma protein responding to inflammation and is used for clinical purposes to diagnose an inflammatory state. For rapid analysis and specific interactions in immunoassays, extensive studies using microfluidic chips have been carried out. Recently, a simple technique to fabricate a disposable PT microchip by a direct printing process was developed and several applications were introduced. One major drawback of the PT microchip, however, is the poor separation performance due to the quality of the microfluidic structures. This problem for a PT microchip can be overcome using a cleavable tag immunoassay, which requires minimal separation performance. After analytes are conjugated onto antibodies which are immobilized on the surface of microbeads placed on the PT microchip, a second group of fluorescently tagged antibodies are added and complexed with the analytes. The tag is then cleaved and the solution containing the cleaved tag is analyzed by electrophoresis. The time needed for the complete analysis to be carried out on a PT microchip was less than 35 min. The dynamic range of the CRP in 10-fold diluted serum was 0.3-100 mg/L and the limit of detection was 0.3 mg/L, which demonstrated the possibility of a quantitative analysis of CRP in serum in clinical trials.
使用聚酯调色剂 (PT) 微芯片进行了用于检测 C 反应蛋白 (CRP) 的芯片上免疫分析。CRP 是一种高度保守的血浆蛋白,对炎症有反应,用于临床诊断炎症状态。为了在免疫分析中进行快速分析和特定相互作用,已经进行了广泛的微流控芯片研究。最近,开发了一种通过直接打印工艺制造一次性 PT 微芯片的简单技术,并介绍了几种应用。然而,PT 微芯片的一个主要缺点是由于微流道结构的质量较差,分离性能不佳。对于 PT 微芯片,这个问题可以通过使用需要最小分离性能的可裂解标签免疫分析来克服。在分析物与固定在放置在 PT 微芯片上的微珠表面上的抗体缀合之后,添加第二组荧光标记的抗体并与分析物复合。然后裂解标签,并通过电泳分析包含裂解标签的溶液。在 PT 微芯片上进行完整分析所需的时间少于 35 分钟。在 10 倍稀释的血清中 CRP 的动态范围为 0.3-100mg/L,检测限为 0.3mg/L,这表明在临床试验中对血清中 CRP 进行定量分析的可能性。