THBS2特异性单克隆抗体的制备与鉴定  

Generation and characterization of THBS2-specific monoclonal antibodies

作　　者：王冰 肖克源 孟麟[1] 寿成超[1] WANG Bing;XIAO Ke-Yuan;MENG Lin;SHOU Cheng-Chao(Key Laboratory of Carcinogenesis and Translational Research,Department of Biochemistry and Molecular Biology,Peking University Cancer Hospital&Institute,Beijing 100142,China)

机构地区：[1]北京大学临床肿瘤学院,北京大学肿瘤医院暨北京市肿瘤防治研究所,生物化学与分子生物学研究室,恶性肿瘤发病机制及转化研究教育部重点实验室,北京100142 [2]山西省长治市人民医院肿瘤内科,长治046000

出　　处：《中国免疫学杂志》2020年第5期588-593,共6页Chinese Journal of Immunology

摘　　要：目的:制备THBS2特异性单克隆抗体并鉴定其特性,为临床应用奠定基础。方法:表达并纯化THBS2不同截断体片段的GST融合蛋白,经小鼠免疫、细胞融合及筛选,制备THBS2特异性单克隆抗体,然后对获得的单抗亲和力和特异性及在不同免疫学实验中的应用进行鉴定,最后采用免疫沉淀法对36例胰腺癌患者及20例正常人血清THBS2进行检测。结果:获得了10株杂交瘤细胞株,均能稳定分泌抗体;10株单抗经ELISA、Western blot和免疫沉淀证实均能特异性识别人THBS2,且发现THBS2在胰腺癌患者血清中的表达水平显著高于正常人(P<0.0001)。结论:成功制备获得了亲和力高、特异性好的抗人THBS2单克隆抗体,为血清THBS2的检测奠定了基础。Objective:To generate and characterize THBS2-specific monoclonal antibodies,and to lay the foundation for clinical applications.Methods:GST fusion proteins of two forms of truncated THBS2 were expressed and purified;monoclonal antibodies against THBS2 were generated by mouse immunization,cell fusion,screening;the specificity and affinity of antibodies were characterized in different immunological experiments;THBS2 level in serum of 36 patients with pancreatic cancer and 20 normal controls was analyzed by immunoprecipitation.Results:10 hybridoma cell lines capable of stably secreting anti-THBS2 antibodies were obtained.These 10 monoclonal antibodies specifically recognized human THBS2 protein in ELISA,Western blot and immunoprecipitation assays.The expression level of THBS2 in serum of patients with pancreatic cancer was significantly higher than that of normal subjects.Conclusion:The anti-human THBS2 monoclonal antibodies with high affinity and specificity were successfully generated,thereby providing the basis for the detection of serum THBS2.

关 键 词：THBS2 胰腺癌 单克隆抗体 制备 鉴定 

分 类 号：R735.9[医药卫生—肿瘤]