Development Of An Ultrasensitive Capillary Electrophores

有必要开发非常敏感的微分析技术来测量生物液体和单细胞中的蛋白质。这种分析的一种方法是毛细管电泳（CE），这是一种仅利用纳升材料的强大分析工具。然而，传统的检测系统不足以分析非常小的生物样品和单细胞，其中许多分析物以微克和飞克的数量存在。激光诱导荧光（LIF）的使用已被证明能够克服许多传统检测系统的缺点。在DBEPS，我们设计并开发了一种实验室建造的LIF检测器，能够测量大约100飞图的荧光色标记分析物。虽然需要进一步改进以确保在分析前对多种分析物进行统一标记，但对更先进的光子探测器的研究正在进行中。目前，一个完整的实验室构建的CE-LIF系统已经建成，能够常规测量0.5 pg/ml水平的分析物。进一步的发展将使这种灵敏度提高到1飞图/毫升或更高的水平。一旦实现这一目标，该仪器将能够可靠地测量临床和研究样品中的单细胞分泌物和/或细胞质。将CE与免疫亲和预分析步骤耦合将进一步完善该仪器的能力，使其能够分析10-100 pg范围内的样品。由于与加拿大的商业微加工设施的互动安排，进一步开发将电泳系统移动到芯片格式已经取得进展。这导致了一种基于芯片的免疫亲和系统的发展，未来将发展为设计和制造一种高度敏感的“芯片实验室”。用于临床分析。

There is a need for the development of very sensitive micro-analytical techniques for measuring proteins in biological fluids and single cells. One approach to such analysis is capillary electrophoresis (CE), which is a powerful analytical tool utilizing only nanoliters of materials. However, conventional detection systems are inadequate for the analysis of very small biological samples and single cells where many analytes are present in pico- and femto-gram quantities. The use of laser-induced fluorescence (LIF) has been demonstrated to be capable of overcoming many of the shortcomings of more conventional detection systems. At DBEPS, we have designed and developed a laboratory-built LIF detector capable of measuring fluorochrome-labeled analytes at approximately 100 femtograms. Although further refinement is required to ensure uniform labeling of multiple analytes pre-analysis, research into more advanced photon detectors is underway. Presently, a complete laboratory-built CE-LIF system has been constructed and is capable of routinely measuring analytes at the 0.5 pg/ml level. Further development will increase this sensitivity to the 1 femtogram/ml level or beyond. Once this is achieved, the instrument will be capable of reliable measurements of single cell secretions and/or cytosol in clinical and research samples. Coupling CE with an immunoaffinity pre-analysis step will further refine the capabilities of this instrument enabling it to analysis samples in the 10-100 pg range. Further development into moving the electrophoresis system into a chip format has progressed due to a interactive arrangement with a commercial micro-fabrication facility in Canada. This has resulted in developments into a chip-based immunoaffinity system with future developments into the design and manufacture of a highly sensitive ?lab-on-a-chip? for clinical analyses.