Department of Medical Biotechnology, Faculty of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Department of Medical Biotechnology, Faculty of Allied Medical Sciences, Iran University of Medical Sciences, Tehran, Iran.
Biotechnol Adv. 2020 Dec;45:107653. doi: 10.1016/j.biotechadv.2020.107653. Epub 2020 Nov 4.
Tag-assisted protein purification is a method of choice for both academic researches and large-scale industrial demands. Application of the purification tags in the protein production process can help to save time and cost, but the design and application of tagged fusion proteins are challenging. An appropriate tagging strategy must provide sufficient expression yield and high purity for the final protein products while preserving their native structure and function. Thanks to the recent advances in the bioinformatics and emergence of high-throughput techniques (e.g. SEREX), many new tags are introduced to the market. A variety of interfering and non-interfering tags have currently broadened their application scope beyond the traditional use as a simple purification tool. They can take part in many biochemical and analytical features and act as solubility and protein expression enhancers, probe tracker for online visualization, detectors of post-translational modifications, and carrier-driven tags. Given the variability and growing number of the purification tags, here we reviewed the protein- and peptide-structured purification tags used in the affinity, ion-exchange, reverse phase, and immobilized metal ion affinity chromatographies. We highlighted the demand for purification tags in the pharmaceutical industry and discussed the impact of self-cleavable tags, aggregating tags, and nanotechnology on both the column-based and column-free purification techniques.
标签辅助蛋白纯化是学术研究和大规模工业需求的首选方法。在蛋白质生产过程中应用纯化标签可以帮助节省时间和成本，但标记融合蛋白的设计和应用具有挑战性。合适的标记策略必须为最终的蛋白质产品提供足够的表达产量和高纯度，同时保持其天然结构和功能。由于生物信息学的最新进展和高通量技术（例如 SEREX）的出现，许多新的标签被引入市场。各种干扰和非干扰标签目前已经将其应用范围扩大到传统的简单纯化工具之外。它们可以参与许多生化和分析特性，并作为可溶性和蛋白质表达增强剂、在线可视化的探针追踪器、翻译后修饰的检测器以及载体驱动的标签。鉴于纯化标签的可变性和不断增加的数量，我们在这里回顾了亲和层析、离子交换层析、反相层析和固定化金属离子亲和层析中使用的蛋白质和肽结构的纯化标签。我们强调了在制药行业对纯化标签的需求，并讨论了自切割标签、聚集标签和纳米技术对基于柱和无柱纯化技术的影响。
