摘要
目的 制备Tropic180 8基因重组蛋白的单克隆抗体 ,并对其生物学特性进行鉴定。方法 用Tropic180 8基因重组蛋白作为抗原免疫BALB/C小鼠 ,取小鼠脾细胞与小鼠骨髓瘤 (SP2 / 0 )细胞融合 ,经ELISA筛选和有限稀释法获得分泌单克隆抗体的细胞株 ,WesternBlot等方法对其生物学特性进行鉴定。结果 获得 2株识别Tropic180 8基因重组蛋白的单克隆抗体的细胞株Ⅱ 4B、Ⅲ 4C。WesternBlot法显示该抗体特异性地识别Tropic180 8基因重组蛋白 ;ELISA法测定杂交瘤细胞培养上清及体内成瘤后产生的腹水的抗体效价分别为 1∶80、 1∶6 0 0 ;杂交瘤细胞染色体数平均为 90～ 10 0 ;亚类鉴定单抗的重链为小鼠IgG1,轻链为κ型。结论 成功地制备了Tropic180 8基因重组蛋白的单克隆抗体 ,为进一步研究Tropic180 8基因重组蛋白的功能提供了良好的基础。
? Objective To prepare the monoclonal antibodies(McAbs)against Tropic 1808 recombinant proteinsand to identify the biological properties. Method SP2/0 myeloma cells were fused with the splenic cells from BALB/Cmicewhich had been immunized intraperitoneally with Tropic 1808 recombinant proteins. After ELISA screening and subcloning positive hybridom cell strains were obtained, and the biological properties of the McAbs were identified by Western Blotting. Result Western blotting demonstrated that two McAbs, designated as Ⅱ4B and Ⅲ4C, could specifically combine with Tropic 1808 recombinant proteins. The titers of the specific antibody in culture supernatant and ascites fluid were found by ELISA to be 1∶80 and 1∶600 respectively. The chromosomal numbers of hybridoma cells were 90～100. The McAbs were IgG1 kappa isotypes. Conclusion McAbs against Tropic 1808 recombinant protein have been established successfully, which has provided a basis for researching the function of the Tropic 1808 recombinant protein. 〔
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2002年第4期458-461,518,共5页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家杰出青年科学基金资助项目(No.39425006)
