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 antigen sTn and Preparation of Monoclonal Antibodies against sTn

作　　者：章万忠 陈智周[2] 范振符[2] 

机构地区：[1]华北石油井下医院检验科,河北任丘062552 [2]中国医学科学院肿瘤医院核医学科,北京100021

出　　处：《标记免疫分析与临床》2007年第4期226-229,共4页Labeled Immunoassays and Clinical Medicine

摘　　要：提纯肿瘤相关糖抗原sTn并研制针对它的单克隆抗体(mAb)。取羊颌下腺经溴棕三甲铵沉淀,经Sepha- rose CL-4B等获取羊颌下腺粘蛋白(OSM),将OSM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。最终获得2株可分泌sTn mAb细胞株(2F6,7E4),均为小鼠IgGl亚类,2株mAb抗相同抗原表位。免疫组化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。本文成功提纯了sTn抗原并制备了2株杂交瘤细胞株,在胃癌的诊断和治疗中有潜在的应用价值。To purify tumor-associated antigen sTn and prepare monoclonal antibody against it, ovine submaxillary mucin（OSM） after precipitation by C19 H42NBr was purified and concentrated by a Sepharose CL-4B column. 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 wasused to screen positive cells and determine the titre of the antibodies. Double antibody competitive inhibition experiment was used to identify the sTn epitope. Immuno-histochemical analysis was employed to locate sTn＇s expression in gastric carcinoma tissues. The results showed that two strains of monoclonal antibodies （2F6 and 7E4） were obtained. Both of them belong to the IgG1 subclass and target to sTn epitope in OSM. Immuno-histochemical analyses showed that sTn were widely expressed in gastric carcinoma tissues, with a positive rate of 71.4%. The high positive rates of sTn are closely related to the stages of the disease. The results suggested that two strains of monoclonal antibodies against sTn were successfully prepared; they could possibly be used in the diagnosis and treatment of gastric cancer.

关 键 词：羊颌下腺粘蛋白 唾液酸化Tn 单克隆抗体 免疫组织化学 

分 类 号：R730.4[医药卫生—肿瘤]