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    Assembly and screening of highly complex and fully human antibody repertoire in yeast
    2.
    发明申请
    Assembly and screening of highly complex and fully human antibody repertoire in yeast 审中-公开
    组装和筛选酵母中高度复杂和完全的人体抗体谱

    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。

    High throughput generation and screening of fully human antibody repertoire in yeast
    5.
    发明申请
    High throughput generation and screening of fully human antibody repertoire in yeast 审中-公开
    在酵母中高产量生成和筛选完全人抗体谱

    公开(公告)号：US20050142562A1

    公开(公告)日：2005-06-30

    申请号：US10792682

    申请日：2004-03-02

    申请人： Li Zhu ,  Shaobing Hua

    发明人： Li Zhu ,  Shaobing Hua

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

    CPC分类号： C07K16/00 ,  C07K2317/21 ,  C07K2317/622

    摘要： Compostions, 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 proteins against a target protein or peptide comprises: expressing a library of tester proteins in yeast cells, each tester protein being a fusion protein comprised of 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： 提供了组合物，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蛋白文库的方法包括：在酵母细胞中表达测试蛋白的文库，每个测试蛋白是由序列在文库内变化的第一多肽亚基组成的融合蛋白，第二多肽 亚单位，其序列在文库内独立于第一多肽而变化，连接肽连接第一和第二多肽亚基; 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。

    High throughput generation and affinity maturation of humanized antibody
    8.
    发明申请
    High throughput generation and affinity maturation of humanized antibody 有权
    人源化抗体的高通量产生和亲和力成熟

    公开(公告)号：US20060246515A1

    公开(公告)日：2006-11-02

    申请号：US11480037

    申请日：2006-06-29

    申请人： Li Zhu ,  Shuanghong Wei ,  Shaobing Hua

    发明人： Li Zhu ,  Shuanghong Wei ,  Shaobing Hua

    IPC分类号： C40B30/06 ,  C40B40/10 ,  C12N1/18

    CPC分类号： C12N15/1041 ,  C12N15/1037 ,  C12N15/1062 ,  C12N15/1086 ,  G01N33/6845 ,  G01N33/6854

    摘要： 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.

    摘要翻译： 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。

    Human monoclonal antibodies against human CXCR4
    9.
    发明授权
    Human monoclonal antibodies against human CXCR4 有权
    针对人CXCR4的人单克隆抗体

    公开(公告)号：US07138496B2

    公开(公告)日：2006-11-21

    申请号：US10360828

    申请日：2003-02-07

    申请人： Shaobing Hua ,  Michelle Haynes Pauling ,  Li Zhu

    发明人： Shaobing Hua ,  Michelle Haynes Pauling ,  Li Zhu

    IPC分类号： C07K16/00 ,  C12P21/08

    CPC分类号： C07K16/2866 ,  A61K39/39541 ,  A61K2039/505 ,  C07K16/00 ,  C07K2317/21 ,  C07K2317/622 ,  C07K2317/76 ,  C07K2319/00 ,  A61K2300/00

    摘要： Compositions are provided that comprise antibody against membrane proteins such as chemokine receptors. In particular, monoclonal human antibodies against human CXCR4 are provided that are capable of inhibiting HIV infection and chemotaxis in human breast cancer cells. The antibodies can be used as prophylactics or therapeutics to prevent and treat HIV infection and cancer, for screening drugs, and for diagnosing diseases or conditions associated with interactions with chemokine receptors.

    摘要翻译： 提供了包含针对膜蛋白如趋化因子受体的抗体的组合物。 特别地，提供能够抑制人乳腺癌细胞中的HIV感染和趋化性的针对人CXCR4的单克隆抗体。 抗体可用作预防或治疗以预防和治疗HIV感染和癌症，用于筛选药物，以及用于诊断与趋化因子受体相互作用相关的疾病或病症。

    Methods and compositions for transferring DNA sequence information among vectors
    10.
    发明授权
    Methods and compositions for transferring DNA sequence information among vectors 失效
    用于在载体之间转移DNA序列信息的方法和组合物

    公开(公告)号：US5989872A

    公开(公告)日：1999-11-23

    申请号：US909525

    申请日：1997-08-12

    申请人： Ying Luo ,  Shaobing Hua ,  Li Zhu

    发明人： Ying Luo ,  Shaobing Hua ,  Li Zhu

    IPC分类号： C12N15/81 ,  C12P19/34 ,  C07H21/04 ,  C12N15/64 ,  C12N15/85

    CPC分类号： C12N15/81

    摘要： Methods and compositions are provided for transferring DNA sequence information from a first vector to a second vector. In the subject methods, a first vector comprising a region of DNA having a sequence of interest is contacted with a set of three pairs of oligonucleotide primers under conditions sufficient to produce three different PCR products, where each product corresponds to a different reading frame. The oligonucleotide primers comprise a first region of sequence identity with the first vector and a second region permissive of site specific recombination with the second vector. The resultant PCR products are combined with the second vector under conditions sufficient for site specific recombination to occur. Also provided are kits for use in performing the subject methods.

    摘要翻译： 提供了将DNA序列信息从第一载体转移到第二载体的方法和组合物。 在本主题方法中，将包含具有感兴趣序列的DNA区域的第一载体与一组三对寡核苷酸引物在足以产生三种不同PCR产物的条件下接触，其中每个产物对应于不同的阅读框。 寡核苷酸引物包含与第一载体具有序列同一性的第一区域和允许与第二载体进行位点特异性重组的第二区域。 将得到的PCR产物与足以进行位点特异性重组的条件与第二载体组合。 还提供了用于执行受试方法的试剂盒。