抗马干扰素-γ单克隆抗体的异硫氰酸荧光素标记及应用  

Preparation and Application of Equine IFN-γ Monoclonal Antibody Labeled with FITC

作　　者：白宇[1] 张维军[1] 童铁钢[1] 王群[1] 徐树兰[1] 孙庆歌[1] 杨涛[1] 吴东来[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《中国生物制品学杂志》2009年第2期188-191,共4页Chinese Journal of Biologicals

基　　金：国家"863"计划(2006AA10A203);黑龙江省自然科学基金重点项目(ZJN-0602-01)

摘　　要：目的用异硫氰酸荧光素(FITC)标记抗马干扰素-γ(IFN-γ)单克隆抗体,为建立马IFN-γ细胞内细胞因子染色方法提供实验材料。方法利用FITC标记纯化的抗马IFN-γ单克隆抗体(SB10),荧光抗体经系列稀释后,对马外周血单核细胞(PBMC)进行细胞内细胞因子IFN-γ单染色的流式细胞术检测,同时与商品化FITC标记的抗牛IFN-γ单克隆抗体(CC302)进行流式细胞术检测对比试验。结果FITC-CC302染色马PBMC,1μl荧光抗体染色106个细胞时,分泌IFN-γ的细胞频率为9.35%,而FITC-SB10经系列稀释染色,在用0.25μl荧光抗体染色106个细胞时,分泌IFN-γ的细胞频率为9.90%。结论所制备的FITC标记的抗马IFN-γ单克隆抗体,可为建立马细胞免疫学研究方法奠定物质基础,进而为建立监测马机体免疫状态和研究机体免疫机制提供技术平台。Objective To label equine IFN-γ monoclonal antibody（McAb） with fluorescein isothioeyanate（FITC） and provide an experimental material for intracellular eytokine staining （ICS） assay of IFN-γ in equine lymphocytes. Methods Purified equine IFN-γ MeAb SB10 was labeled with FITC, then diluted serially and used for ICS assay of IFN-γ in equine PBMCs by flow eytometry. The result was compared with that of flow cytometry by using commercial FITC-labeled bovine IFN-γ MeAb CC302. Results When 106 PBMCs were stained with 1 μl of FITC-CC302, the frequency of PBMCs secreting IFN-γwas 9. 35%. However, when 106 PBMCs were stained with 0. 25 μl of FITC-SB10, the frequency was 9. 90%. Conclusion The prepared FITC-labeled equine IFN-γ McAb laid a material foundation of developing the methods for immunological study on equine cells and provided a technical platform for monitoring the immune state and investigating the immunological mechanism.

关 键 词：马干扰素-γ 单克隆抗体 异疏氰酸荧光素 细胞内细胞因子染色 

分 类 号：Q785[生物学—分子生物学] Q789