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1. 发明申请

WO0200729A3 GENERATION OF LIBRARIES OF ANTIBODIES IN YEAST AND USES THEREOF 审中-公开
标题翻译：抗生素在日本的生成及其用途
公开(公告)号：WO0200729A3
公开(公告)日：2002-06-13
申请号：PCT/US0120542
申请日：2001-06-25
申请人： GENETASTIX CORP , ZHU LI , HUA SHAOBING BENJAMIN
发明人： ZHU LI , HUA SHAOBING BENJAMIN
IPC分类号： C07K16/00 , C12N15/10 , C12Q1/68
CPC分类号： C07K16/00 , C07K2317/21 , C07K2317/622 , C07K2319/80 , C12N15/1055
摘要： Compositions, kits and methods are provided for generating highly diverse libraries of proteins such as antibodies via homologous recombination in vivo , and screening these libraries against protein, peptide and nucleic acid targets using a two-hybrid method in yeast. The method for screening a library of tester fusion proteins against a target protein or peptide comprises: expressing a library of tester proteins in yeast cells, the tester fusion protein comprising a first polypeptide subunit whose sequence varies within the library, a second polypeptide subunit whose sequence varies within the library independently of the first polypeptide, and a linker peptide which links the first and second polypeptide subunits; expressing one or more target fusion proteins in the yeast cells expressing the tester proteins, each of the target fusion proteins comprising a target peptide or protein; and selecting those yeast cells in which a reporter gene is expressed, the expression of the reporter gene being activated by binding of the tester fusion protein to the target fusion protein.
摘要翻译：提供组合物，试剂盒和方法用于通过体内同源重组产生高度多样化的蛋白质文库，例如抗体，并使用酵母中的双杂交方法筛选这些文库对蛋白质，肽和核酸靶标。用于筛选针对靶蛋白或肽的测试融合蛋白文库的方法包括：在酵母细胞中表达测试蛋白的文库，所述测试者融合蛋白包含序列在文库内变化的第一多肽亚基，第二多肽亚基，其序列在文库内独立于第一多肽而变化，以及连接第一和第二多肽亚基的连接肽; 在表达测试蛋白的酵母细胞中表达一种或多种靶融合蛋白，每种靶融合蛋白包含靶肽或蛋白质; 并选择其中表达报告基因的酵母细胞，通过测试融合蛋白与靶融合蛋白的结合来激活报告基因的表达。

2. 发明申请

WO02055718A3 ASSEMBLY AND SCREENING OF HIGHLY COMPLEX AND FULLY HUMAN ANTIBODY REPERTOIRE IN YEAST 审中-公开
标题翻译：组合和筛选高复合物和完全人类抗体代谢物在YEAST
公开(公告)号：WO02055718A3
公开(公告)日：2003-07-24
申请号：PCT/US0151044
申请日：2001-10-31
申请人： GENETASTIX CORP , ZHU LI , HUA SHAOBING BENJAMIN , SHERIDAN JAMES , LIN YU-HUEI
发明人： ZHU LI , HUA SHAOBING BENJAMIN , SHERIDAN JAMES , LIN YU-HUEI
IPC分类号： C07K16/00 , C12N1/19 , C12Q1/68 , C40B30/04 , G01N33/68 , C12N15/81 , C12N15/70 , C12N15/85 , C12N15/86 , G01N33/50
CPC分类号： C40B30/04 , C07K16/00 , C07K2317/21 , C07K2317/55 , C12Q1/6897 , G01N33/6845 , C12Q2565/201
摘要： Compositions, methods, and kits are provided for efficiently generating and screening a library of highly diverse protein complexes for their ability to bind to other proteins or oligonucleotide sequences. In one aspect of the invention, a library of expression vectors is provided for expressing the library of protein complexes. The library comprises a first nucleotide sequence encoding a first polypeptide subunit; and a second nucleotide sequence encoding a second polypeptide subunit. The first and second nucleotide sequences each independently varies within the library of expression vectors. In addition, the first and second polypeptide subunit are expressed as separate proteins which self-assemble to form a protein complex, such as a double-chain antibody fragment (dcFv or Fab) and a fully assembled antibody, in cells into which the library of expression vectors are introduced. The library of expression vectors can be efficiently generated in yeast cells through homologous recombination; and the encoded proteins complexes with high binding affinity to their target molecule can be selected by high throughput screening in vivo or in vitro.
摘要翻译：提供了组合物，方法和试剂盒，用于有效地产生和筛选高度多样化的蛋白质复合物的文库以获得与其它蛋白质或寡核苷酸序列结合的能力。在本发明的一个方面，提供表达载体文库用于表达蛋白质复合物的文库。该文库包含编码第一多肽亚基的第一核苷酸序列; 和编码第二多肽亚基的第二核苷酸序列。第一和第二核苷酸序列各自独立地在表达载体的文库内变化。此外，第一和第二多肽亚基表达为分离的蛋白，其自组装形成蛋白复合物，例如双链抗体片段（dcFv或Fab）和完全组装的抗体，其中文库引入表达载体。表达载体文库可以通过同源重组在酵母细胞中有效产生; 并且可以通过体内或体外的高通量筛选来选择对其靶分子具有高结合亲和力的编码蛋白复合物。

3. 发明专利

AU7155301A High throughput generation and screening of fully human antibody repertoire in yeast 未知
公开(公告)号：AU7155301A
公开(公告)日：2002-01-08
申请号：AU7155301
申请日：2001-06-25
申请人： GENETASTIX CORP
发明人： ZHU LI , HUA SHAOBING BENJAMIN
IPC分类号： C07K16/00 , C12N15/10 , C12Q1/68

4. 发明专利

CA2411515A1 GENERATION OF LIBRARIES OF ANTIBODIES IN YEAST AND USES THEREOF 未知
公开(公告)号：CA2411515A1
公开(公告)日：2002-01-03
申请号：CA2411515
申请日：2001-06-25
申请人： GENETASTIX CORP
发明人： ZHU LI , HUA SHAOBING BENJAMIN
IPC分类号： C07K16/00 , C12N15/10 , C12Q1/68

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