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首页> 外文会议>ASME Internatinal Mechanical Engineering Congress and Exposition >A MATHEMATICAL MODEL OF THE IMMOBILIZED METAL AFFINITY CHROMATOGRAPHY FOR PROTEIN SEPARATIONS: SIMULATION OF LOADING AND ELUTION CONDITIONS
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A MATHEMATICAL MODEL OF THE IMMOBILIZED METAL AFFINITY CHROMATOGRAPHY FOR PROTEIN SEPARATIONS: SIMULATION OF LOADING AND ELUTION CONDITIONS

机译：固定化金属亲和色谱法的数学模型进行蛋白质分离：载荷和洗脱条件的模拟

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Use of numerous steps in purification of proteins which are present in a very low level in a cell extract, lead to a very low yield and high cost. The lack of abundance of such proteins impede studies and applications in immunology, vaccine developments, pharmaceuticals and diagnostics. We have utilized Immobilized Metal Affinity Chromatography (IMAC) to assess the possibility of one step purification protocol for such proteins. The different binding affinities proteins have for immobilized chelated metal ions is the basis for protein fractionation by IMAC. The binding event, in turn, results in the formation of a protein-metal complex, which can easily be reversed by lowering the pH leading to the elution of the protein. When the prerequisites for IMAC retention are absent, genetic modifications may endow the protein with increased metal binding affinity by its fusion to a metal binding peptide (i.e. a tail). Among advantages of IMAC are that the ligand is inexpensive and the affinity matrix can be easily regenerated. We have made significant progress in formulating a mathematical model that incorporates the effects of kinetics of protein-metal binding events, film mass transfer and effective diffusivity of proteins, flowrate, density of the metal binding groups etc. The partial differential equations comprising the model are solved by the spectral method. The model simulates the breakthrough curve and the pH dependent elution of proteins. Moreover, a systematic analysis of performance parameters, reveals optimum operational conditions of loading and elution for attaining high resolution separation of a target protein from a mixture. Experimental work for verification of the model predictions are underway.
机译：在细胞提取物中纯化蛋白质纯化的许多步骤，导致了非常低的产率和高成本。缺乏丰富的这种蛋白质阻碍了免疫学，疫苗发育，药物和诊断中的研究和应用。我们利用固定化的金属亲和层析（IMAC）来评估这种蛋白质的一步纯化方案的可能性。不同的结合亲和力蛋白质具有固定化的螯合金属离子是IMAC蛋白质分级的基础。反过来，结合事件导致形成蛋白质 - 金属络合物，通过降低导致蛋白质洗脱的pH可以容易地逆转。当不存在IMac保留的先决条件时，遗传修饰可以通过其融合到金属结合肽（即尾部）来赋予蛋白质增加的金属结合亲和力。 IMAC的优点是配体廉价，亲和力基质可以容易地再生。在制定蛋白质 - 金属结合事件，膜传质和蛋白质的有效扩散率，流量，金属结合组的有效扩散性等中的作用方面取得了重大进展。包括该模型的部分微分方程是通过光谱法解决了。该模型模拟了蛋白质的突破性曲线和pH依赖性洗脱。此外，对性能参数的系统分析，揭示了用于从混合物中获得高分辨率分离靶蛋白的高分辨率分离的加载和洗脱的最佳运行条件。正在进行验证模型预测的实验工作。

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