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采用自制的多价单抗酶结合物和Dot-ELISA法检测血吸虫病人和感染家兔治疗前后血清循环抗原。报告如下: 1 材料和方法 1.1 单克隆抗体的制备感染血吸虫尾蚴或SEA免疫的BALB/c小鼠脾细胞分别与SP2/O骨髓瘤细胞融合,经克隆化后,用IFA筛选出2株分泌抗血吸虫成虫肠相关阳极抗原(CAA)单克隆抗体杂交瘤细胞株3B_(11)E_3、3B_(10)G_8和1株抗血吸虫卵糖蛋白抗原单克隆抗体杂交瘤细胞株3G_4A_4。将细胞株分别注入小鼠腹腔,诱生的腹水置低温保存。 1.2 多价单克隆抗体酶结合物的制备三种单抗腹水经PEG法提纯后,按相同比例混合成多价单抗,用戊二醛二步法交联辣根过氧化物酶。Dot-ELISA测定酶结合物最适工作浓度为1:1000用于
Self-made multivalent monoclonal antibody conjugates and Dot-ELISA method were used to detect serum circulating antigens in schistosomiasis patients and infected rabbits before and after treatment. The report is as follows: 1 Materials and methods 1.1 Preparation of monoclonal antibodies BALB / c mouse spleen cells infected with Schistosoma japonicum or SEA-immunized mice were fused with SP2 / O myeloma cells respectively. After cloning, Schistosoma japonicum adult gut-associated anodic antigen (CAA) monoclonal antibody hybridoma cell line 3B_ (11) E_3,3B_ (10) G_8 and one anti-schistosome egg glycoprotein antigen monoclonal antibody hybridoma cell line 3G_4A_4. The cell lines were injected into the abdominal cavity of mice, induced ascites stored at low temperature. 1.2 Preparation of polyvalent monoclonal antibody enzyme conjugate Three monoclonal antibodies ascites purified by PEG method, the same ratio mixed multi-monoclonal antibody, two-step glutaraldehyde cross-linked horseradish peroxidase. The optimal concentration of enzyme conjugate for Dot-ELISA was 1: 1000