Generation of human chromosome-specific monoclonal antibodies using trans-chromosomic (TC) mice

使用转染色体 (TC) 小鼠生成人类染色体特异性单克隆抗体

基本信息

  • 批准号:
    13357004
  • 负责人:
  • 金额:
    $ 22.38万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
  • 财政年份:
    2001
  • 资助国家:
    日本
  • 起止时间:
    2001 至 2003
  • 项目状态:
    已结题

项目摘要

TC mice that possess the human chromosome fragments (hCFs) containing the entire human immunoglobulin heavy chain locus and the kappa light chain locus generate fully human monoclonal antibody. Because the TC mice are engineered to neither express endogenous immunoglobulin heavy chain nor kappa light chain, we have generated the anti-human melanoma IgM monoclonal antibody from TC mice that immunize microcell-hybrid mouse melanoma cells, B16-F10 containing a human chromosome 6 as antigen. Furthermore, we also generated a human cell surface molecule specific monoclonal IgG2a antibody from mouse immunized by B16-F10 hybrid cells containing a human chromosome 3. This antibody has a specificity to human cell lines, not to mouse cell lines. Using mass spectrometric analyses, we detected the antigen of this antibody as human Trasferrin Receptor (hTfR), which are located in chromosome 3. In addition, we examined the generation of monoclonal antibody against human cell surface antigens utilizing in vitro differentiated TC-ES 8dTC-ES) cells as immunogens. In a test case, we attempted to generate monoclonal antibody against human neural progenitor cell (NPC) antigen(s) by using the chemically defined medium (CDM) culture for dTC-ES cells containing a human chromosome 4. As a result, we isolated B6-13 antibody that responses to specifically recognize the surface of this cell line. The staining profiles of dTC-ES cells and human embryonic carcinoma (EC) cells with B6-13 were similar to the expression profile of nestin, a well characterized intracellular marker for NPCs. We also identified that B6-13 antigen is CD133 on 4p15.32 using mass spectrometric analyses. Thus, these results suggest that this system for the isolation of a specific human antibody by immunization of malignant or ES cells containing a human chromosome is a valuable resource for drugs and understanding the mechanism that regulates the differentiation in the development.
具有含有整个人免疫球蛋白重链基因座和κ轻链基因座的人染色体片段(hCF)的TC小鼠产生完全人源单克隆抗体。由于TC小鼠经工程改造既不表达内源性免疫球蛋白重链也不表达κ轻链,因此我们从TC小鼠中产生了抗人黑素瘤IgM单克隆抗体,其免疫微细胞杂交小鼠黑素瘤细胞,B16-F10,其含有人6号染色体作为抗原。此外,我们还从含有人3号染色体的B16-F10杂交细胞免疫的小鼠中产生了人细胞表面分子特异性单克隆IgG 2a抗体。该抗体对人细胞系具有特异性,而对小鼠细胞系不具有特异性。使用质谱分析,我们检测到该抗体的抗原为人转铁蛋白受体(hTfR),其位于3号染色体上。此外,我们研究了利用体外分化的TC-ES细胞作为免疫原产生抗人细胞表面抗原的单克隆抗体。在一个测试案例中,我们试图通过使用含有人4号染色体的dTC-ES细胞的化学成分确定的培养基(CDM)培养物来产生针对人神经祖细胞(NPC)抗原的单克隆抗体。结果,我们分离出了B6-13抗体,其响应特异性识别该细胞系的表面。用B6-13对dTC-ES细胞和人胚胎癌(EC)细胞的染色谱与nestin的表达谱相似,nestin是NPC的良好表征的细胞内标记物。我们还利用质谱分析鉴定了B6-13抗原是4p15.32上的CD 133。因此,这些结果表明,通过免疫含有人染色体的恶性细胞或ES细胞来分离特异性人抗体的该系统对于药物和理解调节发育中的分化的机制是有价值的资源。

项目成果

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OSHIMURA Mitsuo其他文献

OSHIMURA Mitsuo的其他文献

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{{ truncateString('OSHIMURA Mitsuo', 18)}}的其他基金

The elucidation of the carcinogenic mechanism of the Down's syndrome using chromosome engineering technology
利用染色体工程技术阐明唐氏综合症的致癌机制
  • 批准号:
    25221308
  • 财政年份:
    2013
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for Scientific Research (S)
Gene therapy of Duchenne muscular dystrophy using own stem cells and human artificial chromosome
使用自身干细胞和人类人工染色体对杜氏肌营养不良症进行基因治疗
  • 批准号:
    21249022
  • 财政年份:
    2009
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Construction of human artificial chromosome for gene therapy of Duchenne muscular dystrophy
杜氏肌营养不良症基因治疗人类人工染色体的构建
  • 批准号:
    18390107
  • 财政年份:
    2006
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Genome-wide analysis of genomic impinting in cancer
癌症基因组印记的全基因组分析
  • 批准号:
    14026029
  • 财政年份:
    2002
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for Scientific Research on Priority Areas
Generation of ES cells that stably its translocation to mouse chromosome
产生稳定易位至小鼠染色体的 ES 细胞
  • 批准号:
    12672200
  • 财政年份:
    2000
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
グリオーマにおけるテロメレース活性およびテロメア長の臨床応用への検討
胶质瘤端粒酶活性和端粒长度检测的临床应用
  • 批准号:
    08457366
  • 财政年份:
    1996
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Chromosomal mupping of a gene responsible for Bloom syndrome via microcell-mediated chromosome Transfer
通过微细胞介导的染色体转移对导致布卢姆综合征的基因进行染色体修饰
  • 批准号:
    04454539
  • 财政年份:
    1992
  • 资助金额:
    $ 22.38万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
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