抗人甲状腺球蛋白单克隆抗体的制备  

Preparation of Monoclonal Antibody against Human Thyroglobulin

作　　者：靳晓瑞 吴意[2] 廖旻晶[1] 邱义兰[3] 唐娜[1] 曾林秀 王平 郭向荣[3] 刘如石[1] JIN Xiaorui;WU Yi;LIAO Minjing;QIU Yilan;TANG Na;ZENG Linxiu;WAN Ping;GUO Xiangrong;LIU Rushi(Hunan Normal University,1.School of Medicine;First Affiliated Hospital;School of Life Science,Changsha 410013,Hunan,China)

机构地区：[1]湖南师范大学医学院,湖南长沙410013 [2]湖南师范大学第一附属医院,湖南长沙410013 [3]湖南师范大学生命科学学院,湖南长沙410013

出　　处：《激光生物学报》2018年第2期166-174,182,共10页Acta Laser Biology Sinica

基　　金：省自然科学基金(2018JJ2226);湖南省重点研发计划(2015SK20332)

摘　　要：目的:制备分泌特异性抗人甲状腺球蛋白(thyroglobulin,Tg)单克隆抗体的杂交瘤细胞株,为建立高灵敏度的Tg检测方法做准备。方法:以天然人源甲状腺球蛋白为抗原经皮下免疫BALB/c小鼠,通过细胞融合制备分泌抗人甲状腺球蛋白单克隆抗体,并对其进行特异性鉴定,建立检测Tg的双抗体ELISA(enzyme-linked immunosorbent assay)夹心法。结果:获得7株可稳定分泌抗人甲状腺球蛋白单克隆抗体的杂交瘤细胞株,经ELISA鉴定,筛选抗体可与Tg抗原有良好的特异性反应。建立的双抗体夹心ELISA方法敏感性可达1 ng/mL。结论:成功制备了抗人Tg单克隆抗体并建立了检测人Tg双抗体夹心ELISA方法,为进一步研发Tg快速诊断试剂盒提供了原料。Objective： To prepare monoclonal antibody against human thyroglobulin and establish a highly sensitive method to detect human thyroglobulin. Methods： BALB/c mice were subcutaneously immunized by native human thyroglobulin,the splenocytes of immunized mice were fused with SP2/0 myeloma cells by a routine method. The hybridoma cells secreting specific antibody were detected. The stability of the obtained hybridoma cells and the specificity of antithyroglobulin monoclonal antibody the hybridoma cells secreted were identified. Based on all above,the double antibody sandwich ELISA method was established to detect thyroglobulin. Results： Seven hybridoma cell lines which could secrete monoclonal antibody stably were obtained,and ELISA indicated the antibodies had a good specificity and sensitivity to Tg antigen. A sandwich ELISA kit for detecting human thyroglobulin was established,and its sensitivity reached to 1 ng/m L. Conclusion： The anti-thyroglobulin monoclonal antibodies were prepared and a sandwich ELISA kit for the detection of human thyroglobulin was established successfully,which would provide a raw material for the development of Tg rapid diagnostic kits.

关 键 词：单克隆抗体 甲状腺球蛋白 纯化 酶联免疫吸附实验 

分 类 号：R392[医药卫生—免疫学]