Advanced IMAC Methodology

固定化金属离子亲和层析(IMAC)已被证明是从复杂混合物中分离微量蛋白质的有效手段。该奖项的目的是表征和绘制几种IMA吸附剂，并利用这些研究的结果来制定和测试使用级联模式多亲和层析(CASMAC)从粗品混合物中分离蛋白质的方案。第一个目标是从螯合剂、金属离子和层析条件等方面对新型凝胶进行表征。在其他方法中，将使用吸附等温线和核磁共振。其次，这些凝胶将通过氨基酸、模型肽和蛋白质的保留研究来绘制。第三，研究人员计划开发和测试允许使用CASMAC对复杂蛋白质混合物进行分级的协议。预计这项工作将导致向科学界提供IMAC和CASMAC的吸附剂，这些吸附剂为蛋白质和多肽提供递增的亲和力，并专门针对不同的氨基酸侧基相互作用。此外，研究人员计划开发一种合理的方法，使用一系列亲和技术来有效分离复杂的蛋白质混合物。

Immobilized metal ion affinity chromatography (IMAC) has proved useful in isolating proteins present in trace amounts from complex mixtures. The objective of this award is to characterize and map several IMA-adsorbents and to use the results of these studies to formulate and test protocols for the isolation of proteins from crude mixtures using cascade mode multi-affinity chromatography (CASMAC). The first goal is to characterize new gels with regard to chelating agents, metal ions and chromatography conditions. Among other methods, adsorption isotherms and NMR will be used. Secondly, these gels will be mapped using retention studies of amino acids, and model peptides and proteins. Thirdly, the investigators plan to develop and test protocols that allow fractionation of complex protein mixtures using CASMAC. It is anticipated that this work will result in making adsorbents available to the scientific community for both IMAC and CASMAC that offer incremental affinities for proteins and peptides, and that are specifically targeted for interaction with different amino acid side groups. Further the investigators plan to develop a rational approach using a series of affinity techniques to separate complex protein mixtures effectively.