原核表达的PD-1与PD-L1融合蛋白相互作用分析  

Analyze the bioactivity of PD-1 and PD-L1 recombination protein which expressed by prokaryotic system

作　　者：沈立萍[1] 许龙[2] 李建东[1] 陈斯勇[1] 郭瑜[1] 邱丰[1] 毕胜利[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京100052 [2]中国军事医学科学院生物工程研究所

出　　处：《中华实验和临床病毒学杂志》2009年第6期424-426,共3页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的分析经原核表达纯化的PD-1和PD-L1融合蛋白分子间相互作用力及其生物学活性。方法PD-1蛋白经过预浓缩后，用氨耦联的原理固定于CM5芯片表面，用HBS-EP缓冲液梯度稀释PD-L1蛋白，以20μl/min流速注射到耦联PD-1的传感芯片通道上，结合3min，解离15min；观察两者结合情况并用BIA Evaluation软件4进行分析。结果PD-1在缓冲液的pH值为4.5、浓度为40μg／ml的状态下能稳定耦联于CM5芯片表面，Ru值约为3300；稀释度为200mmol／ml的PD-Ll与PD-1能较好地结合，RU值为150，亲和常数为KD=3.5×10^-6。结论经原核表达纯化的PD-1与PD-L1融合蛋白具有较强的特异性亲和力及良好的生物学活性，为下一步研究的开展打下基础。Objective To analyze the interaction of PD-1 and PD-L1 recombination protein and to know their bioactivity and affinity. Methods Stick the PD-1 protein on the surface of CM5 sensor chip by the method of Ammine coupling after being preconsentrated. Dilute the PD-L1 protein step by step and reject it to the passage on CM5 sensor chip which had been stick by PD-1. The time of combination is 3 minutes and of separation is 15 minutes, respectively. Observe the procession and analyze data by BIA Evaluation software 4. Results On the consistency of 40μg/ml, pH 4.5, the PD-1 protein could couple steady on the surface of CM5sensor chip, RU is 3300. On the density of 200 mmol/ml PD-L1 could combine with PD-1 specifically, RU = 150,KD = 3.5×10^-6 . Conclusion The PD-1 and PD-L1 recombination protein which we expressed by prokaryotic system have good affinity and bioactivity. The results could provide basic condition for later study.

关 键 词：病毒融合蛋白质类 膜蛋白质类 受体 免疫 基因表达调控 病毒 生物学鉴定法 

分 类 号：R341[医药卫生—基础医学]