Optimization of IMAC by Fine-Tuning of Protein Metal Interactions

9713286 Ataai目标蛋白的纯化是其生产总成本的主要部分。纯化的高成本往往限制了这种蛋白质的可用性，这反过来又阻碍了免疫学研究、疫苗开发以及用于人类保健的药物和诊断试剂的生产。尽管免疫亲和纯化应该允许纯化这些靶蛋白，但它存在许多缺点，包括配体的高成本，并且在某些情况下，靶蛋白与抗体的强结合，这可能需要使用极端的pH值或其他恶劣条件来洗脱蛋白质。IMAC在纯化融合到金属亲和性尾部的蛋白质方面显示出了希望。基于初步的结果，组合肽库的筛选可以在一个色谱步骤中确定纯化目标融合蛋白的肽，回收率高，纯度高，成本低。＊＊＊

9713286 Ataai Purification of a target protein constitutes the major fraction of the total cost of its production. The high cost of purification often limits the availability of such proteins, which in turn impedes studies in immunology, vaccine development, and the production of pharmaceutical and diagnostic reagents for human health care. Although immunoaffinity purification should allow for purification of such target proteins, it suffers from a number of shortcomings including high cost of the ligand, and, in some cases, strong binding of the target protein to the antibody, which could require the use of extremes of pH or other harsh conditions for the elution of the protein. IMAC has shown promise for the purification of proteins fused to metal affinity tails. Based on preliminary results, screening of combinatorial peptide libraries may identify peptides capable of purifying target fusion proteins in one chromatography step with high recovery, purity, and low cost. ***