【摘要】 目的 :研制抗rhNDPK A (recombinanthumannucleosidediphosphatekinase A)单克隆抗体 (mAb) ,并鉴定其特性。 方法 :以纯化的rhNDPK A免疫BALB/c小鼠 ,采用杂交瘤技术制备抗rhNDPK AmAb ;用免疫双扩散鉴定Ig亚类 ;West ernblot鉴定mAb的特异性 ;间接ELISA检测mAb的腹水效价、亲和常数 ,并进行表位分析。结果 :获得 6株可分泌特异性mAb的抗rhNDPK A的杂交瘤细胞系 2D9、8C7、13E2、15D9、15E3和 2 0D9,Ig亚类均为IgG1;其效价为 1× 10 -4～5× 10 -6;亲和常数为 4 .5× 10 -9～ 2 .8× 10 -10 mol/L ;共有3个抗原表位。结论 :获得抗rhNDPK A的mAb ,为进一步用于临床诊断和实验研究创造了条件更多还原

【Abstract】 AIM: To prepare monoclonal antibodies (mAb) against recombinant human nucleoside diphosphate kinase-A(NDPK-A) and characterize their properties. METHODS: BALB/c mice were immunized with rhNDPK-A, and mAb was prepared by hybridoma technique. Ig subclass and specificity was analysed by double immunodiffusion and western blot respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA. RESULTS: 6 hybridoma cell lines secreting anti-rhNDPK-A mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 6 mAbs in ascitic fluid were 1×10 -4-5×10 -6. Relative affinity of mAbs were 4.5×10 -9-2.8×10 -10 mol/L. They recognized 3 different epitopes on rhNDPK-A molecule. CONCLUSION: 6 mAbs against rhNDPK-A have been prepared successfully which provide useful reagent for clinical diagnosis and further research.更多还原