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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
Objective: To develop a monoclonal antibody (mAb) against rhNDPK A and identify its characteristics. Methods: BALB / c mice were immunized with purified rhNDPK A and rhNDPK AmAb was prepared by hybridoma technique. The Ig subtype was identified by double immunodiffusion. The specificity of mAb was identified by West ernblot. The indirect ELISA was used to detect the ascites titer of mAb. And constants, and epitope analysis. RESULTS: Six anti-rhNDPK A secreting mAbs were obtained from hybridoma cell lines 2D9, 8C7, 13E2, 15D9, 15E3 and 20D9. The Ig subtypes were IgG1 and their titer was 1 × 10 -4 ~ 5 × 10 -6; the affinity constant is 4.5 × 10 -9 ~ 2 .8 × 10 -10 mol / L; there are 3 antigenic epitopes. Conclusion: The mAb against rhNDPK A can be used for further clinical diagnosis and experimental research