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    • 62. 发明申请
    • High throughput generation and affinity maturation of humanized antibody
    • 人源化抗体的高通量产生和亲和力成熟
    • US20060246515A1
    • 2006-11-02
    • US11480037
    • 2006-06-29
    • Li ZhuShuanghong WeiShaobing Hua
    • Li ZhuShuanghong WeiShaobing Hua
    • C40B30/06C40B40/10C12N1/18
    • C12N15/1041C12N15/1037C12N15/1062C12N15/1086G01N33/6845G01N33/6854
    • Compositions, methods, and kits are provided for efficiently generating and screening humanized antibody with high affinity against a specific antigen. The library of humanized antibody is generated by mutagenizing a chimeric antibody template that combines human antibody framework and antigen binding sites of a non-human antibody. Alternatively, the library of humanized antibody is generated by grafting essential antigen-recognition segment(s) such as CDRs of the non-human antibody into the corresponding position(s) of each member of a human antibody library. This library of humanized antibody is then screened for high affinity binding toward a specific antigen in vivo in organism such as yeast or in vitro using techniques such as ribosome display or mRNA display. The overall process can be efficiently performed in a high throughput and automated manner, thus mimicking the natural process of antibody affinity maturation.
    • 提供了组合物,方法和试剂盒,用于高效地产生和筛选对特异性抗原具有高亲和力的人源化抗体。 人源化抗体文库通过诱变组合人抗体框架和非人抗体的抗原结合位点的嵌合抗体模板产生。 或者,人源化抗体文库通过将必需的抗原识别片段(例如非人抗体的CDR)接枝到人抗体文库的每个成员的对应位置来产生。 然后在生物体如酵母或体外使用诸如核糖体展示或mRNA显示的技术,筛选人源化抗体文库以体内对体内特异性抗原的高亲和力结合。 整个过程可以以高通量和自动化方式有效地进行,从而模仿抗体亲和力成熟的自然过程。
    • 63. 发明授权
    • Assembly and screening of highly complex and fully human antibody repertoire in yeast
    • 组装和筛选酵母中高度复杂和完全的人体抗体谱
    • US06610472B1
    • 2003-08-26
    • US09703399
    • 2000-10-31
    • Li ZhuShaobing Benjamin HuaJames SheridanYu-Huei Lin
    • Li ZhuShaobing Benjamin HuaJames SheridanYu-Huei Lin
    • G01N3353
    • C40B30/04C07K16/00C07K2317/21C07K2317/55C12Q1/6897G01N33/6845C12Q2565/201
    • 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.
    • 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。