抗人PD-1高亲和力抗血清的制备及其在PD-1免疫印迹分析中的应用  

Preparation of anti-human PD-1 antiserum with high-affinity and its application in PD-1 immunoblotting analysis

作　　者：施焕敬[1] 徐丽慧[2] 韩捷[1] 卢宏松[1] 何贤辉[1] 

机构地区：[1]暨南大学生命科学技术学院组织移植与免疫实验中心,广东广州510632 [2]暨南大学生命科学技术学院生物工程研究所,广东广州510632

出　　处：《暨南大学学报（自然科学与医学版）》2008年第3期315-319,共5页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金项目(30572199、30230350、30371651);生物化学与分子生物学广东省重点学科资助()

摘　　要：利用纯化的可溶性人PD-1胞外域(sPD-1)蛋白为免疫原免疫小鼠,获得抗血清.免疫印迹显示该抗血清与原核表达的sPD-1有特异性反应,但与某些菌体蛋白存在交叉反应.经菌体蛋白吸附法处理后,交叉反应明显减弱.以商业化抗人PD-1抗体和小鼠sPD-1抗血清分析经亲和层析富集的活化的Jurkat T细胞表达的PD-1,只有自制的抗血清鉴定到特异反应条带,其表观分子质量为49 ku.The anti-human soluble program death-1 （sPD-1） antiserum was prepared by immunizing mice with the purified recombinant sPD-1. Immunoblotting analysis showed that this antiserum had specific reaction with sPD-1, and had some cross-reactivity with bacterial proteins. This cross-reactivity was significantly decreased after treating the antiserum with whole bacterial lysates. PD-1 expressed by activated Jurkat T cells was enriched by affinity chromatography, and assayed by commercial antibody and the antiserum respectively. Only the antiserum identified the natural PD-1, its apparent molecular weight is about 49 000.

关 键 词：程序死亡蛋白-1 JURKAT T细胞 抗血清 免疫印迹 

分 类 号：R392.12[医药卫生—免疫学]