EGFRvⅢ单抗的制备及其特异性的研究  

Preparation of EGFRvⅢ monoclonal antibody and identification of its characterization

作　　者：许泼实[1] 韩双印[2] 魏威[3] 魏晓霞[1] 万震[1] 

机构地区：[1]河南省人民医院检验科,河南郑州450003 [2]河南省人民医院中心实验室 [3]南方医科大学生物技术学院

出　　处：《现代预防医学》2012年第17期4500-4501,4503,共3页Modern Preventive Medicine

基　　金：河南省自然科学基金(072102310021)

摘　　要：目的制备EGFRvⅢ单克隆抗体,并对其生物学特性进行鉴定。方法将小鼠骨髓瘤细胞NS-1与免疫的BALB/C小鼠,免疫小鼠脾细胞进行融合,通过酶联免疫吸附试验(ELISA)筛选出EGFRvⅢ阳性的杂交瘤细胞株,单抗与EGFRvⅢ抗原结合的特性性通过Western blot进行鉴定。建立裸鼠人肺细胞癌和表皮细胞癌移植瘤模型,分别腹腔注射EGFRvⅢ单抗,尼妥珠单抗(Nimotuzumab),生理盐水,观察3组别的抑瘤效果和作用。结果用单克隆抗体技术筛选得到的杂交瘤细胞株而制备的EGFRvⅢ单抗,经过Western blot鉴定,结果为可以特异性识别EGFRvⅢ。EGFRvⅢ单抗,尼妥珠单抗对人表皮细胞癌移植瘤的抑瘤率分别为84%和86.3%。结论通过杂交瘤技术制备的EGFR vⅢ单克隆抗体,特异性好,在抑制肿瘤方面效果显著,对进一步的机制研究和临床应用有一定价值。OBJECTIVE To prepare the EGFRvⅢ monoclonal antibody,and analyze the bionomics.METHODS Myeloma NS-1 cells and spleen cells of BALB/c mice were mixed together,and the positive anti-EGFRvⅢ monoclonal antibody hybridoma cells were selected and identified by ELISA.The specific binding between antibody and antigen was tested by Western blot.The transplantable tumor mouse models were established.The effects of inhibiting tumor was observed by treating mice with EGFRvⅢ monclonal antibody,Nimotuzumab and physiological saline.RESULTS By the hybridism technique,we got the EGFRvⅢ monoclonal antibody and the binding of the antibody with antigen was specific,which tested by Western blot.The inhibitory rates of EGFRvⅢ monoclonal antibody and Nimotuzumab against A431 transplanted tumors were 84% and 86.3% respectively.CONCLUSION Our study shows that the EGFRvⅢ monoclonal antibody which obtained by hybridoma technique has a good specificity and ability of inhibiting tumor growth.This will be of great value to further investigation in mechanisms and clinical application.

关 键 词：表皮生长因子受体 单克隆抗体 表皮生长因子受体三型突变体 移植瘤 

分 类 号：R730.3[医药卫生—肿瘤]