Optimization of IMAC by Fine-Tuning of Protein Metal Interactions

通过微调蛋白质金属相互作用来优化 IMAC

• 批准号: 9713286
• 负责人: Mohammad Ataai
• 金额: $ 19.15万
• 依托单位: University of Pittsburgh
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-01 至 2001-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9713286&HistoricalAwards=false
• 关键词: Optimization; IMAC; Fine; Tuning; Protein

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. ***

9713286目标蛋白的提纯占其生产总成本的主要部分。高昂的提纯成本往往限制了这些蛋白质的可获得性，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。虽然免疫亲和纯化可以纯化这些目标蛋白，但它存在一些缺点，包括配基成本高，在某些情况下，目标蛋白与抗体结合很强，这可能需要使用极端的pH或其他苛刻的条件来洗脱蛋白。IMac在纯化融合到金属亲和尾巴上的蛋白质方面表现出了希望。根据初步结果，组合多肽文库的筛选可以在一个层析步骤中鉴定出能够纯化目的融合蛋白的多肽，且回收率高、纯度高、成本低。***