肿瘤相关糖抗原sTn的提纯及其单克隆抗体的制备  

Purification of tumor-associated sTn antigen and establishment of monoclonal antibodies against sTn

作　　者：章万忠 陈智周[2] 范振符[2] 

机构地区：[1]河北省华北石油井下医院检验科,河北任丘062522 [2]中国医学科学院肿瘤医院核医学科,北京100021

出　　处：《国际检验医学杂志》2008年第1期23-26,共4页International Journal of Laboratory Medicine

摘　　要：目的提纯肿瘤相关糖抗原sTn并研制针对它的单克隆抗体（mAb）。方法取羊颌下腺经溴棕三甲铵沉淀，Sepharose CL-4B凝胶过滤等获取羊颌下腺黏蛋白（OSM），将其作为抗原采用脾内注射、完全佐剂2种方法免疫Balb／C小鼠。常规方法细胞融合，用ELISA间接法筛选阳性细胞并测定效价，双抗体竞争抑制试验识别sTn表位，采用免疫组化检测sTn在胃癌组织中的表达。结果获得2株可分泌sTnmAb的杂交瘤细胞株2F6、7E4，免疫球蛋白亚类测定均为小鼠IgG1亚类，2株mAb抗相同抗原表位。免疫组织化学染色显示，胃癌组织中sTn表达阳性率较高，可达71．4％，且与胃癌进程相关。结论成功提纯了sTn抗原并制备了2株针对它的杂交瘤细胞株，在胃癌的诊断和治疗中有潜在的应用价值。Objective To purify tumor-associated sTn antigen and develop monoclonal antibodies against sTn. Methods After precipitatation by cetrimonium bromide, ovine submaxillary mucin （OSM） was concentrated through Sepharose CL-4B column gel filtering. OSM was used as the antigen to immunize Balb/C mice in two different ways： direct spleen injection and the routine immunization. After cell fusion, indirect ELISA was used to screen positive cells and the titre of the antibodies were determined. Double-antibody sandwich assay was used to identify sTn epitope. Immuno histochemical analysis was employed to locate the expression of sTn in gastric carcinoma tissue. Results Two hybridoma cell lines （2F6 and 7E4） secreting sTn mAb were acquired. Both of them belonged to the IgG1 subclass and were targeted to the same sTn epitope in OSM. Immunohistochemical analysis showed sTn was highly expressed in gastric carcinoma tissue, with a positive rate of 71.4%. And the positive rate was closely related to the process of gastric cancer. Conclusion Successfully purification of sTn antigen and establishment of two hybridoma cell lines against sTn processes potential applicable value in the diagnosis and treatment of gastric cancer.

关 键 词：黏蛋白类 抗体 单克隆 免疫组织化学 胃肿瘤 

分 类 号：R730.45[医药卫生—肿瘤] R735.2[医药卫生—临床医学]