Three-in-One Chromatography-Free Purification, Tag Removal, and Site-Specific Modification of Recombinant Fusion Proteins Using Sortase A and Elastin-iike Polypeptides

Joseph J. Bellucci; Miriam Amiram; Jayanta Bhattacharyya

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Three-in-One Chromatography-Free Purification, Tag Removal, and Site-Specific Modification of Recombinant Fusion Proteins Using Sortase A and Elastin-iike Polypeptides

机译：使用分选酶A和弹性蛋白酶样多肽的重组融合蛋白的三合一无色谱纯化，标签去除和位点特异性修饰

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Column chromatography is the workhorse of protein purification, but the requirements for large volumes of buffers, expensive resins that have limited potential for re-use, and significant operator hours to optimize and execute the separations present major financial and technical hurdles to scaling up production. Additionally, covalent modification of proteins with small molecules and polymers is routine, but available conjugation methods frequently produce heterogeneous products as a result of incomplete reactivity or the presence of multiple reaction sites within a protein. These two unit operations currently present major limitations to the production of recombinant proteins at the scales necessary for both research and manufacturing.

机译：柱色谱法是蛋白质纯化的主要手段，但对大量缓冲液，昂贵的树脂（重用潜力有限）以及大量操作工时间以优化和执行分离的要求，对扩大生产规模构成了主要的财务和技术障碍。另外，用小分子和聚合物对蛋白质进行共价修饰是常规操作，但是由于不完全的反应性或蛋白质内多个反应位点的存在，可用的缀合方法经常产生异质产物。目前，这两个单元操作对研究和生产所必需的规模的重组蛋白的生产提出了主要限制。

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《Angewandte Chemie》 |2013年第13期|共6页
作者
Joseph J. Bellucci; Miriam Amiram; Jayanta Bhattacharyya;
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正文语种 eng
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anticancer agents :: bioorganic chemistry :: biotechnology :: protein expression :: Sortase A;

机译：抗癌剂::生物有机化学::生物技术::蛋白质表达:: Sortase A;

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1. Three-in-One Chromatography-Free Purification, Tag Removal, and Site-Specific Modification of Recombinant Fusion Proteins Using Sortase A and Elastin-iike Polypeptides [J] . Joseph J. Bellucci, Miriam Amiram, Jayanta Bhattacharyya Angewandte Chemie . 2013,第13期

机译：使用分选酶A和弹性蛋白酶样多肽的重组融合蛋白的三合一无色谱纯化，标签去除和位点特异性修饰
2. A self-cleavable sortase fusion for one-step purification of free recombinant proteins [J] . Mao HY Protein Expression and Purification . 2004,第1期

机译：一种可自我裂解的分选酶融合蛋白，用于一步纯化游离重组蛋白
3. Sortase-Tag Expressed Protein Ligation: Combining Protein Purification and Site-Specific Bioconjugation into a Single Step [J] . Robert Warden-Rothman, Ilaria Caturegli, Vladimir Popik, Analytical chemistry . 2013,第22期

机译：Sortase-Tag表达的蛋白连接：将蛋白纯化和位点特异性生物结合结合到一个步骤
4. Sortase A-mediated site-specific immobilization of peptides and proteins for interactome analysis by LC-MS/MS [C] . Eberhard Krause, Benno Kuropka, Nadine Royla, International Mass Spectrometry Conference . 2014

机译：分类序列A介导的肽和蛋白质的特异性固定蛋白质，用于通过LC-MS / MS进行蛋白酶分析
5. The CBM9 fusion tag: A new technology for inexpensive production and affinity purification of recombinant proteins [D] . Kavoosi, Mojgan 2007

机译：CBM9融合标签：一种用于廉价生产和亲和纯化重组蛋白的新技术
6. Three-in-one chromatography-free purification tag removal and site-specific modification of recombinant fusion proteins using Sortase A and elastin-like polypeptides [O] . Joseph J. Bellucci, Miriam Amiram, Jayanta Bhattacharyya, -1

机译：三合一色谱 - 自由纯化标签除去以及使用分子酶A和弹性蛋白样多肽的重组融合蛋白的特异性修饰
7. Three-in-One Chromatography-Free Purification, Tag Removal, and Site-Specific Modification of Recombinant Fusion Proteins Using Sortase A and Elastin-like Polypeptides [O] . Joseph J. Bellucci, Miriam Amiram, Jayanta Bhattacharyya, 2013

机译：使用三相色谱 - 无选蛋白 - 无组分纯化，并使用分子酶A和弹性蛋白样多肽的重组融合蛋白的特异性修饰

Three-in-One Chromatography-Free Purification, Tag Removal, and Site-Specific Modification of Recombinant Fusion Proteins Using Sortase A and Elastin-iike Polypeptides

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