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1. 发明申请

US20070065913A1 ISOLATION OF BINDING PROTEINS WITH HIGH AFFINITY TO LIGANDS 有权
标题翻译：分离具有高亲和力的结合蛋白
公开(公告)号：US20070065913A1
公开(公告)日：2007-03-22
申请号：US11461757
申请日：2006-08-01
申请人： Gang Chen , Andrew Hayhurst , Jeffrey Thomas , Brent Iverson , George Georgiou
发明人： Gang Chen , Andrew Hayhurst , Jeffrey Thomas , Brent Iverson , George Georgiou
IPC分类号： C12P21/06 , C07H21/04 , C12N9/00 , C12N1/21 , C07K14/195 , C12N15/74
CPC分类号： C12N15/1034
摘要： The invention overcomes the deficiencies of the prior art by providing a rapid approach for isolating binding proteins capable of binding small molecules and peptides via “display-less” library screening. In the technique, libraries of candidate binding proteins, such as antibody sequences, are expressed in soluble form in the periplasmic space of gram negative bacteria, such as Escherichia coli, and are mixed with a labeled ligand. In clones expressing recombinant polypeptides with affinity for the ligand, the concentration of the labeled ligand bound to the binding protein is increased and allows the cells to be isolated from the rest of the library. Where fluorescent labeling of the target ligand is used, cells may be isolated by fluorescence activated cell sorting (FACS). The approach is more rapid than prior art methods and avoids problems associated with the surface-expression of ligand fusion proteins employed with phage display.
摘要翻译：本发明通过提供通过“无显示”文库筛选分离能够结合小分子和肽的结合蛋白的快速方法来克服现有技术的缺陷。在该技术中，候选结合蛋白（例如抗体序列）的文库以可溶形式表达在革兰氏阴性细菌如大肠杆菌的周质空间中，并与标记的配体混合。在表达对配体具有亲和性的重组多肽的克隆中，与结合蛋白结合的标记配体的浓度增加，并允许细胞与文库的其余部分分离。当使用靶配体的荧光标记时，可以通过荧光激活细胞分选（FACS）分离细胞。该方法比现有技术方法更快速，并避免与噬菌体展示所用配体融合蛋白的表面表达相关的问题。

2. 发明授权

US07871796B2 Isolation of binding proteins with high affinity to ligands 有权
标题翻译：结合蛋白与配体的高亲和力分离
公开(公告)号：US07871796B2
公开(公告)日：2011-01-18
申请号：US11461757
申请日：2006-08-01
申请人： Gang Chen , Andrew Hayhurst , Jeffrey G. Thomas , Brent L. Iverson , George Georgiou
发明人： Gang Chen , Andrew Hayhurst , Jeffrey G. Thomas , Brent L. Iverson , George Georgiou
IPC分类号： C12P21/06 , C07H21/04
CPC分类号： C12N15/1034
摘要： The invention overcomes the deficiencies of the prior art by providing a rapid approach for isolating binding proteins capable of binding small molecules and peptides via “display-less” library screening. In the technique, libraries of candidate binding proteins, such as antibody sequences, are expressed in soluble form in the periplasmic space of gram negative bacteria, such as Escherichia coli, and are mixed with a labeled ligand. In clones expressing recombinant polypeptides with affinity for the ligand, the concentration of the labeled ligand bound to the binding protein is increased and allows the cells to be isolated from the rest of the library. Where fluorescent labeling of the target ligand is used, cells may be isolated by fluorescence activated cell sorting (FACS). The approach is more rapid than prior art methods and avoids problems associated with the surface-expression of ligand fusion proteins employed with phage display.
摘要翻译：本发明通过提供通过“无显示”文库筛选分离能够结合小分子和肽的结合蛋白的快速方法来克服现有技术的缺陷。在该技术中，候选结合蛋白（例如抗体序列）的文库以可溶形式表达在革兰氏阴性细菌如大肠杆菌的周质空间中，并与标记的配体混合。在表达对配体具有亲和性的重组多肽的克隆中，与结合蛋白结合的标记配体的浓度增加，并允许细胞与文库的其余部分分离。当使用靶配体的荧光标记时，可以通过荧光激活细胞分选（FACS）分离细胞。该方法比现有技术方法更快速，并避免与噬菌体展示所用配体融合蛋白的表面表达相关的问题。

3. 发明授权

US07083945B1 Isolation of binding proteins with high affinity to ligands 有权
标题翻译：结合蛋白与配体的高亲和力分离
公开(公告)号：US07083945B1
公开(公告)日：2006-08-01
申请号：US09699023
申请日：2000-10-27
申请人： Gang Chen , Andrew Hayhurst , Jeffrey G. Thomas , Brent L. Iverson , George Georgiou
发明人： Gang Chen , Andrew Hayhurst , Jeffrey G. Thomas , Brent L. Iverson , George Georgiou
IPC分类号： C12P21/06 , C12P21/04 , C12N15/09 , G01N33/53
CPC分类号： C12N15/1034
摘要： The invention overcomes the deficiencies of the prior art by providing a rapid approach for isolating binding proteins capable of binding small molecules and peptides via “display-less” library screening. In the technique, libraries of candidate binding proteins, such as antibody sequences, are expressed in soluble form in the periplasmic space of gram negative bacteria, such as Escherichia coli, and are mixed with a labeled ligand. In clones expressing recombinant polypeptides with affinity for the ligand, the concentration of the labeled ligand bound to the binding protein is increased and allows the cells to be isolated from the rest of the library. Where fluorescent labeling of the target ligand is used, cells may be isolated by fluorescence activated cell sorting (FACS). The approach is more rapid than prior art methods and avoids problems associated with the surface-expression of ligand fusion proteins employed with phage display.
摘要翻译：本发明通过提供通过“无显示”文库筛选分离能够结合小分子和肽的结合蛋白的快速方法来克服现有技术的缺陷。在该技术中，候选结合蛋白（例如抗体序列）的文库以可溶形式表达在革兰氏阴性细菌如大肠杆菌的周质空间中，并与标记的配体混合。在表达对配体具有亲和性的重组多肽的克隆中，与结合蛋白结合的标记配体的浓度增加，并允许细胞与文库的其余部分分离。当使用靶配体的荧光标记时，可以通过荧光激活细胞分选（FACS）分离细胞。该方法比现有技术方法更快速，并避免与噬菌体展示所用配体融合蛋白的表面表达相关的问题。

4. 发明申请

US20140243234A1 NUCLEIC ACID COMPOSITIONS, METHODS AND KITS FOR RAPID PAIRING OF AFFINITY AGENTS 审中-公开
标题翻译：核酸组成物，快速配对亲和剂的方法和工具
公开(公告)号：US20140243234A1
公开(公告)日：2014-08-28
申请号：US14236837
申请日：2012-08-03
申请人： Andrew Hayhurst
发明人： Andrew Hayhurst
IPC分类号： C07K16/46 , G01N33/68 , C12N15/10
CPC分类号： C07K16/46 , C07K2319/00 , C07K2319/21 , C07K2319/90 , C12N15/1037 , C12P21/02 , G01N33/6854
摘要： The present inventions relate to the selection and production of specific proteins or peptides with desired properties. The inventions relate to the agents and methods of identifying select peptides or proteins with specific binding properties or greater enzymatic performance from vast numbers of variants.
摘要翻译：本发明涉及具有所需性质的特异性蛋白质或肽的选择和生产。本发明涉及鉴定具有特异性结合特性的选择肽或蛋白质的代理和方法，或者来自广泛变体的更高的酶性能。

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