Development of an application to quantify clinically relevant metalloproteins in human blood plasma********

可以很容易地获得血浆并对其进行蛋白质分析，以获得有关个人健康状况的有用诊断信息。与血浆分析固有的困难是，它含有数千种蛋白质。然而，如果能够对金属蛋白进行分析，这个问题就可以大大简化，金属蛋白代表含有结合金属的蛋白质的一个子组。因此，开发一种可以用来确定血浆中所有含金属蛋白质的方法需要分离它们，并利用适当的金属特定检测器进行检测。为此，Gailer博士的团队开发了一种基于尺寸排除色谱(SEC)的分离方法，用于直接分析血浆中含有的铁、铜和锌金属蛋白，这些蛋白通过电感耦合等离子体原子发射光谱仪(ICP-AES)进行检测。这种方法使用30厘米×1.0厘米秒的色谱柱，每次分析需要0.5毫升，需要25分钟，这对临床生化实验室来说不是很有吸引力。这种方法的小型化将大大减少分析所需的等离子体数量，并缩短整体分析时间。分离可以用较小的SEC柱(15 cm x 0.5 cm)完成，含铁蛋白质可以通过石墨炉原子吸收光谱仪(GFAAS)检测。由于Lumex仪器公司生产GFAAS仪器，盖勒博士将开发一种SEC-GFAAS系统，用于快速分析人体血浆中临床相关的铁金属蛋白。这种联用技术有可能为这家加拿大仪器制造商打开新的市场，因为拟议的SEC-GFAAS系统能够快速和专门地定量人体血浆中的含铁蛋白，这对临床生物化学实验室很有吸引力。*

Blood plasma can be easily obtained and analyzed for proteins to obtain useful diagnostic information about the health status of an individual. The difficulty that is inherently associated with the analysis of plasma is that it contains thousands of proteins. This problem, however, can be dramatically simplified if it can be analyzed for metalloproteins, which represent a sub-group of proteins that contain bound metals. The development of a method that can be used to determine all metal-containing proteins in plasma therefore requires their separation and their detection by means of appropriate metal-specific detectors. To this end, Dr. Gailer's group has developed a size exclusion chromatography (SEC)-based separation method for the direct analysis of blood plasma for the contained iron, copper and zinc-containing metalloproteins, which are detected by means of an inductively coupled plasma atomic emission spectrometer (ICP-AES). This method employs a 30 cm x 1.0 cm SEC column, requires 0.5 mL/analysis and requires 25 min, which is not attractive for clinical biochemistry labs. The miniaturization of this method would significantly decrease the amount of plasma needed for analysis and reduce the overall analysis time. The separation can be accomplished with a smaller SEC column (15 cm x 0.5 cm) and the iron-containing proteins may be detected by means of a graphite furnace atomic absorption spectrometer (GFAAS). Since Lumex Instruments manufactures a GFAAS instrument, Dr. Gailer will develop a SEC-GFAAS system for the rapid analysis of human blood plasma for clinically relevant iron metalloproteins. This hyphenated technique has the potential to open new markets for this Canadian instrument manufacturer since capability of the proposed SEC-GFAAS system to rapidly and specifically quantify iron-containing proteins in human plasma is attractive for clinical biochemistry laboratories. ********************************