摘要
【目的】构建1个人源性抗人肺癌单链抗体库，并对抗体库进行筛选。【方法】用肺鳞癌病人肺门淋巴结来提取总RNA，通过噬菌体展示技术构建人源性单链抗体库，用人肺癌细胞系（A549）和人正常肺组织细胞初筛及鉴定抗体库。【结果】构建了1个库容量为3．1×10^7噬菌体单链抗体库，抗体库克隆的表达产物可与人肺癌细胞结合，筛选出1个可与人肺癌细胞系A549相对特异性结合的噬菌体单链抗体，此单链抗体与A549结合的浓度比人正常肺组织细胞低约130倍，其基因片段大小约为750bp，其重链和轻链基因片段约370bp。【结论】本研究成功构建了1个人源性抗人肺癌单链抗体库，并筛选出1个与肺癌细胞系A549相对特异性单链抗体。
[Objective] To construct and screen a library of human single antibody against human lung cancer. [Methods]Total RNA was extracted from metastatic lymph node of lung hilum of a patient with lung squamocellular cancer. A library of human single antibody against human lung cancer was constructed by phage display technique, screened primarily and identified with A549 lines, lung cancer cells, and human normal lung cells. [Results] A antibody library, containing 3.1 × 10^7 clones, was constructed successfully. The antibody expressed by clones of the antibody library could react with A549 lines. A relative specific page single antibody against A549 lines was panned successfully. The lowest concentration for it to react with human normal lung cells was about 130 times as high as that with A549 lines. The gene sequences of single chain antibody, high chain and light chain were respectively about 750bp, 370bp and 370bp. [Conclusion] A library of human single antibody against human lung cancer is constructed successfully. A relative specific page single antibody against A549 lines is panned successfully.
出处
《医学临床研究》
CAS
2009年第10期1780-1783,1787,共5页
Journal of Clinical Research
基金
广州市卫生局科研基金（2005-YB-109）
关键词
肺肿瘤
抗体
肿瘤
lung neoplasms
antibodies,neoplasm
