抗人内皮细胞生长抑制素单克隆抗体的制备和鉴定

Preparation and Identification of Monoclonal Antibodies Against Human Endostatin

研制抗内皮细胞生长抑制素(Endostatin,ES)单克隆抗体,为ES制剂提供更有效的质量控制手段和为内皮细胞抑制素介入基因治疗肿瘤提供临床评价工具。将重组的人ES蛋白纯化,免疫BALB/c小鼠,取ES蛋白免疫的小鼠脾细胞与小鼠骨髓细胞Ag8.653进行细胞融合,对杂交瘤细胞进行筛选,阳性孔经3次有限稀释法亚克隆,获得4株鼠抗人ES蛋白的杂交瘤细胞株,分别命名为:1C2、6C4、6B4、6C5,以间接ELISA、Western免疫印迹等方法进行抗体鉴定。结果表明4株杂交瘤细胞分泌的单抗具有效价高,特异性好的特点,且免疫球蛋白亚型均为IgG1。其中1C2亲和力强,具有较高的应用价值。所得单克隆体具有一定的特异性和敏感性,可为ES的深入研究提供更好的条件。

In order to obtain mouse anti-human ES monoclonal antibodies for potent approaches to quality control of ES preparations and ES mediated cancer gene therapy, BALB/c mice were immunized with human ES recombinant protein SDS-PAGE gel suspension. Spleen cells were fused with AgS. 653 myeloma cells. The fused cells were selected by HAT medium and screened by ELISA. The monoclonal antibodies were identified by ELISA and Western blotting. Four clones of hybrdoma stable in secreting ES mAbs were obained. The antibodies showed the high specificity to ES with strong affinity. ES monoclonal antibodies produced could bind specifically to ES protein providing a valid approach in ES research and identification.