摘要
目的制备抗人胰腺癌细胞单克隆抗体(McAb),并对其特性进行鉴定。方法以胰腺癌细胞系Patu8988免疫Balb／ c小鼠,取其脾细胞与小鼠骨髓瘤细胞SP2／0进行融合,经选择培养,筛选及克隆化后,建立杂交瘤细胞系。以ELISA flow cytometry等方法研究抗体特性。以MTT法研究细胞毒作用。结果得到一株能够稳定分泌特异性抗体的杂交瘤细胞系 A7。用琼脂糖双扩散鉴定其为IgG1亚类,ELISA测定其腹水效价为:1:16000。流式细胞仪检测表明:该抗体与Patu8988 细胞呈阳性反应(>90％),与肝癌、结肠癌、胃癌、肺癌、乳腺癌呈阴性反应(<5％)。MTT法示该抗体对胰腺癌细胞的杀伤率明显高于对照组。结论 McAbA7对胰腺癌的诊断和治疗可能具有一定的临床意义。
Objective To prepare and identify monoclonal antibody（McAb） of human pancreatic cancer associated antigen.Methods Balb/c ices were immunized with human pancreatic cancer cell line Patu8988 and the lymphocytes hybridoma technique was used for McAb preparation.The property of antibody was studied with ELISA,MTT, flow cytometry methods.Results One hybridoma cell line （A7）,which secretes monoclonal antibody against pancreatic carcinoma, was established and its titer in ascites was 1：1.6×10^4 Flow cytometry showed that Mab A7 was positive with pancreatic cancer cells,but did not react with liver cancer, colon,cancer, lung cancer, gastric cancr, breast cancer cell.MTT assay proved that the cytotoxicitis of effective groups were higher than control groups.Mab was typed by reaction with subclass-specific （rabbit） anti-mouse antibody and was found to be IgG 1.Conclusion MabA7 with high immuno-reactivity and tissue specificity may perhaps be useful for diagnosis and treatment of pancreatic cancer.
出处
《中国血液流变学杂志》
CAS
2006年第1期20-22,共3页
Chinese Journal of Hemorheology
关键词
胰腺癌
单克隆抗体
制备
鉴定
pancreatic cancer
monoclonal antibody
preparation
identification
