期刊文献+

PCR扩增抗人视网膜母细胞瘤单克隆抗体轻链可变区基因

Amplifying Variable Region Gene of Light Chain of Monoclonal Antibody against Human Retinoblastoma by PCR
下载PDF
导出
摘要 目的:获得抗人视网膜母细胞瘤单克隆抗体轻链可变区基因。方法:从分泌抗人视网膜母细胞瘤单克隆抗体的杂交瘤细胞中提取总RNA,逆转录形成cDNA,用扩增小鼠K轻链可变区基因的引物,通过RT-PCR方法,扩增基因,1.5%琼脂糖凝胶电泳鉴定。结果:用轻链引物扩增获得340bp的基因片段。结论:从三株杂交瘤细胞中均获得高纯度的总RNA;用RT-PCR方法从一株杂交瘤细胞成功克隆了抗人视网膜母细胞瘤单克隆抗体轻链可变区基因,为基因工程抗体研究奠定了良好基础。眼科学报2001;17:194~197。 Objective: To acquire the variable region gene of light chain of monoclonal antibodyagainst human retinoblastoma.Methods: Total RNA were extracted from hybridoma cells secreting specific monoclonalantibody (MeAb) against human retinoblastoma (RB), then transcripted reversely intocDNA with random primers. The variable region of the light chain (VL) gene fragmentswere ampliflied using polymerase chain reaction (PCR) method. Agrose gelelectrophoresis was confirmed.Results: 1.5% agrose gel electrophoresis indicated that VL. gene was about 340 basepairs.Conclusion: The light chain variable region gene of the McAb against human RB wasamplified successfully, which lays a good basis for construction of a recombinantantibody. Eye Science 2001; 17: 194 ~ 197.
出处 《眼科学报》 2001年第4期194-197,共4页 Eye Science
基金 国家自然科学基金(资助号:39870801) 广东省自然科学基金(资助号:98011) 中山医科大学"211"工程项目(资助号:98007)资助
关键词 单克隆抗体 视网膜母细胞瘤 可变区基因 PCR monoclonal antibody, retinoblastoma, variable region gene
  • 引文网络
  • 相关文献

参考文献4

  • 1[5]Eshhar Z, Waks T, Gross G, et al: Specific activiation and targeting of cytotoxic lymphocytes through chimeric single chains consisting of antibody-binding domains and the γ or ξ subunits of the immunoglobulin and T-cell receptors.Proc. Natl. Acad. USA 1993;90: 720.
  • 2[7]Colapinto EV, Humphrey PA, Zalutskky MR, et al:Comparative localization of murine monoclonal antibody mel14 F(ab 2 fragment and whole IgG2a in human glllioma xenografts. Cancer Res 1988 ;48:5701.
  • 3[8]Char DH, Wood IS, Huhta K, et al: Retinoblastoma: tissue culture lines and monoclonal antibody studies. IOVS 1984;25: 30.
  • 4[10]Orlandi R, Gussow DH, Jones PT, et al: Cloning immunoglobulin variable domains for expression by the polymerase chain reaction. Proc. Natl. Acad. Sci. USA 1989; 86: 3833.
;
使用帮助 返回顶部