Development Of An Ultrasensitive Capillary Electrophores

需要开发非常灵敏的微量分析技术来测量生物液和单细胞中的蛋白质。这种分析的一种方法是毛细管电泳法(CE)，这是一种仅使用纳米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，在这些样品中，许多分析物以皮克和毫微克的数量存在。激光诱导荧光(LIF)的使用已被证明能够克服更传统检测系统的许多缺点。在DBEPS，我们设计和开发了一种实验室建造的激光诱导荧光检测器，能够测量大约100毫微克的氟铬标记分析物。尽管需要进一步改进以确保多个分析物在分析前的统一标记，但对更先进的光子探测器的研究正在进行中。目前，一套完整的实验室构建的CE-LIF系统已经建成，能够对0.5pg/ml的分析物进行常规测量。进一步的开发将把这种灵敏度提高到1毫微克/毫升或更高的水平。一旦实现这一点，该仪器将能够可靠地测量临床和研究样本中的单细胞分泌物和/或胞浆。将CE与免疫亲和预分析步骤相结合，将进一步完善该仪器的能力，使其能够分析10-100 pg范围的样品。由于与加拿大的一家商业微型制造设施进行了互动安排，将电泳系统转移到芯片格式的进一步开发取得了进展。这导致了基于芯片的免疫亲和系统的开发，并在未来开发设计和制造高灵敏的芯片上实验室？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.