摘要
目的:制备TREM2特异性单克隆抗体并对其特异性、亲和力及在免疫学检测中的应用进行鉴定,为TREM2的抗体药物研发奠定基础。方法:表达并纯化TREM2胞外段GST融合蛋白,用其作为抗原免疫小鼠,筛选抗血清效价较高的小鼠,经过细胞融合和单克隆筛选,制备TREM2的特异性单克隆抗体。对获得的单克隆抗体的特异性、亲和力及其在免疫学实验中的应用进行检测。结果:获得了29株TREM2单克隆抗体,其中的24株可与TREM2特异性结合;29株单抗与TREM2结合的EC50均在nmol以上;它们可分别在Western blot、免疫沉淀、流式细胞术和免疫荧光中用于对TREM2的特异检测。结论:成功制备并获得了亲和力高、特异性好的抗TREM2单克隆抗体,为TREM2靶点的成药性抗体开发奠定了基础。
Objective:To preparation TREM2-specific monoclonal antibodies and to evaluation their properties and applica-tions lay a foundation for the development of druggable antibodies against TREM2 targets.Methods:The GST fusion protein containing the extracellular segment of TREM2 was expressed and purified,which was used as antigen to immunize mice.The mouse with high antiserum titer was used to generate specific monoclonal antibodies against TREM2 through cell fusion and monoclonal screening.The specificity and affinity of the monoclonal antibodies and their applications in different immunological experiments were detected.Results:A total of 29 strains of anti-TREM2 monoclonal antibodies were obtained,of which 24 strains could specifically bind to TREM2.The EC50 of antibodies were calculated above nanomolar.These monoclonal antibodies could be used for specific detection of TREM2 in Western blot,immunoprecipitation,flow cytometry and immunofluorescence experiments.Conclusion:Monoclonal anti-bodies against TREM2 with high affinity and specificity are successfully generated,which lay the foundation for the research and the development of antibody drugs targeting TREM2.
作者
马宾
孟麟
戎卓娜
赵传科
寿成超
MA Bin;MENG Lin;RONG Zhuona;ZHAO Chuanke;SHOU Chengchao(Department of Biochemistry and Molecular Biology,Peking University College of Clinical Cancer,Peking University Cancer Hospital and Beijing Institute of Cancer Prevention,Key Laboratory of Malignant Tumor Pathogenesis and Transformation Research of Ministry of Education,Peking University,Beijing 100142,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2024年第1期168-173,177,共7页
Chinese Journal of Immunology
关键词
TREM2
单克隆抗体
制备
鉴定
TREM2
Monoclonal antibody
Preparation
Identification
