Tang Lin, Zheng Lu, Qi Chun-jian, Ma Hong-bing, Zhou Xuan, Zhuang Yu-Mei, Luo Xian-fu, Xu Ying, Zhang Xue-guang
Institute of Medical Biotechnology, Key Laboratory of Medicine and Clinical Immunology of Jiangsu Province, Soochow University, Suzhou 215007, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Sep;23(9):841-3.
To prepare a novel functional mouse anti-human CD40 monoclonal mAb.
Female BALB/c mice of 6-8 weeks old were immunized with CD40 transfectant (L929-CD40) as immunogen. The spleen B cells of the mice were fused with Sp2/0. The hybridoma cells were screened with CD40 transfectant (L929-CD40) by FCM. Fast-strip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio/5C11 competitive assay. The proliferation and cell cycle of tumor cells in vitro were studied by MTT assay and PI staining respectively.
One hybridoma cell line named 2B6 was obtained, which had the property of secreting anti-human CD40 monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 molecule and promoted the proliferation of tumor cells in vitro.
One hybridoma cell line which can secret a novel functional mouse anti-human CD40 mAb has been developed successfully. This mAb can specifically recognize human CD40 and influence the growth of tumor cells in vitro.
制备一种新型功能性小鼠抗人CD40单克隆抗体。
以CD40转染细胞（L929-CD40）作为免疫原，免疫6-8周龄雌性BALB/c小鼠。取小鼠脾脏B细胞与Sp2/0细胞进行融合。采用流式细胞术（FCM）用CD40转染细胞（L929-CD40）对杂交瘤细胞进行筛选。进行快速试纸分析以鉴定该单克隆抗体的Ig亚类。通过生物素/5C11竞争试验检测该单克隆抗体识别的表位。分别采用MTT法和PI染色法研究肿瘤细胞体外增殖及细胞周期。
获得一株名为2B6的杂交瘤细胞系，其具有持续稳定分泌抗人CD40单克隆抗体的特性。该单克隆抗体可特异性识别人类CD40分子，并促进肿瘤细胞体外增殖。
成功构建了一株能分泌新型功能性小鼠抗人CD40单克隆抗体的杂交瘤细胞系。该单克隆抗体可特异性识别人类CD40并影响肿瘤细胞体外生长。
