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

血浆可以很容易地获得和分析蛋白质，以获得有关个人健康状况的有用诊断信息。血浆分析固有的困难在于它含有数千种蛋白质。然而，如果可以分析金属蛋白，这个问题就可以大大简化，金属蛋白代表了含有结合金属的蛋白质的一个子组。因此，开发一种可用于测定血浆中所有含金属蛋白质的方法需要通过适当的金属特异性探测器对它们进行分离和检测。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，这些金属蛋白是通过电感耦合等离子体原子发射光谱仪（ICP-AES）检测的。该方法采用30 cm × 1.0 cm的SEC柱，需要0.5 mL/次分析，需要25分钟，这对于临床生物化学实验室来说并不具有吸引力。该方法的小型化将显著减少分析所需的血浆量，并减少总体分析时间。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 Instruments生产GFAAS仪器，Gailer博士将开发SEC-GFAAS系统，用于临床相关的人血浆铁金属蛋白的快速分析。由于SEC-GFAAS系统能够快速、特异性地定量人体血浆中的含铁蛋白，因此这种连字符技术有可能为这家加拿大仪器制造商打开新的市场，这对临床生物化学实验室具有吸引力。********************************

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. ********************************