公开(公告)号：US06610472B1

公开(公告)日：2003-08-26

申请号：US09703399

申请日：2000-10-31

申请人： Li Zhu ,  Shaobing Benjamin Hua ,  James Sheridan ,  Yu-Huei Lin

发明人： Li Zhu ,  Shaobing Benjamin Hua ,  James Sheridan ,  Yu-Huei Lin

IPC分类号： G01N3353

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。

公开(公告)号：US20050123996A1

公开(公告)日：2005-06-09

申请号：US10947610

申请日：2004-09-21

申请人： Li Zhu ,  Shaobing Hua ,  James Sheridan ,  Yu-Huei Lin

发明人： Li Zhu ,  Shaobing Hua ,  James Sheridan ,  Yu-Huei Lin

IPC分类号： C07K16/00 ,  C12N1/19 ,  C12Q1/68 ,  C40B30/04 ,  G01N33/68 ,  G01N33/53 ,  C12N1/18 ,  C12N15/74 ,  G01N33/543

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 vary 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。