抗桥粒斑蛋白Ⅱ单克隆抗体的研制及鉴定  

Preparation and Characterization of Moncclonal Antibodies to Desmoplakin Ⅱ

作　　者：明洪[1] 杨举伦[1] 武云[1] 罗学林[1] 曾卫玲[1] 蔡琳[1] 

机构地区：[1]昆明医学院第一附属医院中心实验室,成都军区昆明总医院病理科

出　　处：《单克隆抗体通讯》1995年第1期44-47,共4页

摘　　要：本文用改良构椽酸－枸椽酸钠法，从新鲜牛鼻表皮制备的ＤｅｓｍｏＰｌａｋｉｎⅡ（ＤＰⅡ）免疫ＢＡＬＢ／Ｃ，小鼠，取其脾淋巴细胞在ＰＥＧ介导下与ＳＰ２／０细胞融合，经筛选、克隆化获得４株能稳定分泌ＤＰⅡＭＣＡｂ的杂交癌细胞系（分别命名为ＤＰｙ—１，２，３，４）。４株ＭｃＡｂ均为ⅠｇＧＩ，染色体数目为９３～１０５．免疫组化分析表明，４株ＤＰⅡＭｃＡｂ具有很高的上皮组织特异性和敏感性，并提示ＤＰⅡ可能不存在于甲状腺滤泡上皮组织中。腺上皮细胞胞浆内ＤＰⅡＭｃＡｂ免疫组化染色阳性，可能是腺上皮细胞胞浆内存在ＤＰⅡ相关抗原的结果。实验表明ＤＰⅡＭｃＡｂ既可用于冰冻切片，又可用于石蜡切片，为上皮源性肿瘤的病理诊断和发生机理研究提供了一种有用的探针．Spleen cells of BALB/c mice immunized with desmoplakin 11 which was prepared fromfresh cow nose epidermis were fused with myeloma cell line Sp2/0.Antibodies in culture supernatantof the hybridoma were detected by indirect ELISA and immunohistochemical analysis and POsitivehybridomas were cloned using liciting dilutions.The linmunoglobulin subclasses of four DP 11McAbs (DPy-1,2,3,4,)were IgGI and the number of chromosomes of 4 hybridoma cell lines rangedfrom 93-105.Immunohistochemical analysis showed all DPy-1,2,3,4 had high speCificity and sensitivity to epithelia and DP Ⅱ may not exist in epithelial cells of thyroid follicles. The POsitive reaction incytoplasm of glandular epithelial cells stained by ABC homunohistocllemical method with DP 11 McAbmay suggest that DP Ⅱ associated antigen existed in cytoplasm of glandular epithelial cells. The resultSalso showed that DP Ⅱ McAb be used not only in frozen section but also in routinly processed paraffinsection.It is believed that DP Ⅱ McAb is a very useful probe for pathological diagnosis and research onmechanism of tumors derived from epithelial tissues.

关 键 词：单克隆抗体 桥粒斑蛋白II 制备 鉴定 

分 类 号：R392－33[医药卫生—免疫学]