【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。更多还原

【Abstract】 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 1.37 μ mol/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.更多还原