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BALB/C 小鼠腹腔经硅胶或液体石蜡处理后,接种分泌抗胎儿甲种球蛋白的单克隆抗体杂交瘤株(10~7细胞).接种后24h 开始向腹腔注射8μci~3H—亮氨酸溶液,每日1次,共5次.接种后第7d 处死小鼠.取腹水、肝、脾和脑组织.腹水经离心后分离为腹腔细胞和上清.上清经DEAE—纤维素离子交换层析分离为单克隆抗体和其他组分.各组织、单克隆抗体及其他组分经适当处理后用液体闪烁仪检测其放射性.结果发现,各组织(细胞)中以杂交瘤细胞(占腹腔细胞的78.5%)的比放射性最高,层析所分离出的蛋白质组分中以单克隆抗体的比放射性最高.
BALB/C mice were treated with silica gel or liquid paraffin in the peritoneal cavity and were inoculated with a monoclonal antibody hybridoma strain (10~7 cells) secreting anti-fetal gamma globulin. Intraperitoneal injection of 8μci~3H-leucine was started 24 h after inoculation. Solution, once daily, 5 times in total. Mice were sacrificed 7 days after inoculation. Ascites fluid, liver, spleen, and brain tissue were collected. Ascites fluid was separated by centrifugation into peritoneal cells and supernatant. The supernatant was deacetylated by DEAE-cellulose. The chromatographic separation was performed on monoclonal antibodies and other components. Each tissue, monoclonal antibody, and other components were treated appropriately and detected for radioactivity using a liquid scintillation counter. The results showed that hybridoma cells (occurring in the abdominal cavity) were present in each tissue (cell). The specific radioactivity of 78.5% of the cells was the highest, and the specific radioactivity of monoclonal antibody was the highest among the protein fractions separated by chromatography.