基于表面等离子共振技术鉴定鸡补体受体2与其配体chC3 d的亲和力  

The Identification of Binding Affinity between Chicken ComplimentReceptor 2 and Its Ligands chC3 d by Surface Plasmon Resonance

作　　者：靳换 杨芊芊 王聪 涂敏 赵蕾[1] 澎湃 王海良 孟昭英 李永清[1] 章振华[1] JIN Huan;YANG Qian-qian;WANG Cong;TU Min;ZHAO Lei;PENG Pai;WANG Hai-liang;MENG Zhao-ying;LI Yong-qing;ZHANG Zhen-hua(Institute of Animal Husbandry and Veterinary Medicine,Beijing Academy of Agriculture and Forestry Sciences.;China Animal Husbandry Industry Co.,Ltd.;Beijing Lingyu Biotechnology Co.,Ltd.;Beijing Veterinary Medicine and Feed Monitoring Center)

机构地区：[1]北京市农林科学院畜牧兽医研究所,北京100097 [2]中牧实业股份有限公司,北京100000 [3]北京翎羽生物科技有限公司,北京101302 [4]北京市兽药饲料监测中心,北京102200

出　　处：《中国兽药杂志》2023年第7期1-7,共7页Chinese Journal of Veterinary Drug

基　　金：国家自然科学基金青年基金(32202793);北京市农林科学院青年基金(QNJJ202111);北京市农林科学院畜牧兽医研究所改革与发展(XMS202313);北京市农林科学院创新能力建设(KJCX20230424)。

摘　　要：鸡补体受体2(Chicken complement receptor 2,chCR2)是一种表达于鸡B淋巴细胞表面的跨膜蛋白,chCR2可以与其生理学配体鸡补体组分3 d(Chicken complement component 3 d,chC3 d)相互作用。目前评估受体与配体亲和力的方法主要有表面等离子共振技术(Surface plasmon resonance,SPR)和生物膜干涉技术(Biolayer interferometry,BLI)等。旨在利用表面等离子共振技术鉴定chCR2受体分子与其配体chC3 d分子的亲和力。成功构建了重组真核表达载体pTT5-chCR2和pTT5-chC3 d;利用HEK 293F表达系统表达了His-chCR2和His-chC3 d蛋白;利用NI柱纯化系统纯化了His-chCR2和His-chC3 d蛋白;利用表面等离子共振技术,通过分析chCR2与chC3 d结合的动力学参数得出两者之间的平衡解离常数KD为1.37μmol/L。结果表明chCR2与其配体chC3 d之间的亲和力较高。Chicken complement receptor 2(chCR2)is a transmembrane protein expressed on the surface of chicken B lymphocytes.chCR2 can interact with its physiological ligand chicken complement component 3 d(chC3 d).At present,the methods to evaluate the affinity between receptor and ligand mainly include Surface plasmon resonance(SPR)and biomembrane interference(BLI).In this study,the SPR was to identify the affinity between the chCR2 and its ligand chC3 d.The recombinant eukaryotic expression vectors pTT5-chCR2 and pTT5-chC3 d were successfully constructed;His-chCR2 and His-chC3 d proteins were expressed using the HEK 293F expression system;The His-chCR2 and His-chC3 d proteins were purified using the NI column purification system;The equilibrium dissociation constant KD between chCR2 and chC3 d is 1.37μmol/L by analyzing the kinetic parameters of the combination of chCR2 and chC3 d.The results indicate that chCR2 binds to its ligand chC3 d with high affinity.

关 键 词：鸡补体受体2 鸡补体组分3 d 表面等离子共振技术 相互作用 

分 类 号：S859.79[农业科学—临床兽医学]