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。与血浆分析固有相关的困难在于它含有数千种蛋白质。然而，这个问题可以大大简化，如果它可以分析金属蛋白，这代表了一个亚组的蛋白质，含有结合金属。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检测器对其进行分离和检测。为此，Gailer博士的研究小组开发了一种基于尺寸排阻色谱（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，这些金属蛋白通过电感耦合等离子体原子发射光谱仪（ICP-AES）进行检测。该方法采用30 cm x 1.0 cm SEC色谱柱，每次分析需要0.5 mL，需要25 min，这对于临床生化实验室没有吸引力。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，Gailler博士将开发一种SEC-GFAAS系统，用于快速分析人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开辟新市场，因为拟议的SEC-GFAAS系统能够快速特异地定量人血浆中的含铁蛋白质，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. ********************************