A two-dimensional (2-D, weak anion exchange chromatography-gel electrophoresis, WAX-GE) separations coupling to inductively coupled plasma mass spectrometry (ICP-MS) strategy was developed for efficient tracking of metalloproteins. Samples were fractionized with the primary dimension (WAX) and the collected fractions were subsequently subjected to the secondary dimension (GE) and finally detected with ICP-MS. The metalloproteins of interest from the secondary dimension were online split and collected for further manipulation. The molecular weight of metalloprotein was calibrated with a home-made protein marker including six iodine-labelled proteins with molecular weights ranging from 14 kDa to 77 kDa. The developed 2-D system is of higher separation resolution as compared to one-dimensional (1-D) system. Metalloproteins in Escherichia coli were further examined for validation of the method preformation. Several known or unknown metal-associated proteins were identified, evidencing the feasibility of the proposed method. Taken together, we established a solid methodology for efficient separation and qualitative study of metalloproteomics, providing a new strategy for the metallomics research.

二维（2-D，弱阴离子交换色谱-凝胶电泳，WAX-GE）分离与电感耦合等离子体质谱法（ICP-MS）的耦合被开发出来，可以有效地跟踪金属蛋白。样品采用一级色谱（WAX）进行分馏，然后将收集的级分进行二级色谱（GE），最后用ICP-MS检测。从次要维度感兴趣的金属蛋白在线拆分并收集以进行进一步处理。金属蛋白的分子量用自制的蛋白质标记物校准，该标记物包括六种碘标记的蛋白质，分子量范围为14 kDa至77 kDa。与一维（1-D）系统相比，开发的2-D系统具有更高的分离分辨率。中的金属蛋白进一步检查大肠杆菌以验证方法的形成。鉴定了几种已知或未知的金属相关蛋白，证明了所提出方法的可行性。综上所述，我们建立了一种可靠的方法，可以有效地分离和定性研究金属组学，为金属组学研究提供了新的策略。