抗钙调素单克隆抗体的制备及鉴定

The Preparation and Evaluation of Monoclonal Antibody against Calmodulin

目的 制备及鉴定抗钙调素单克隆抗体。方法 以碳化二亚胺法将钙调素 卵清蛋白偶联物作为免疫原 ,采用常规免疫方法免疫BALB/C小鼠 ,杂交瘤技术制备抗牛脑钙调素单克隆抗体。酶联免疫吸附法 (ELISA)检测 ,应用硫酸铵盐析法和离子交换层析法进行抗体纯化。分别采用免疫印迹、免疫酶斑点法鉴定抗钙调素单克隆抗体的性质。结果 脾细胞与骨髓瘤细胞的融合率为 85 %。阳性克隆率为 0 .35 % ,获得两株动物钙调素单克隆抗体 ,抗体效价为 1∶10 0 0 ,对钙调素有较强的专一性。结论 此研究制备出了高特异性、高亲和力及高效价的抗牛脑钙调素单克隆抗体。

Objective To study preparation and identification of a monoclonal antibody against calmodulin. Methods The mouse was primaryly immuned by a immunal antigen which was a concation of calmodulin and albumine by EDC assay.Positive clons which were picked out by ELISA were then purified using amine suphate pricipitation followed by ion column chromatography. Results The fusional rate of splenocyte and myeloma cell,positive clonic rate and anitibody valence was 85%,0.35% and 1∶1000 respectively.Two anticalmodulin monoclonal antibodies were obtained. Conclusion The anticalmodulin monoclonal antibody with valence is obtained from this preparation.