心肌肌球蛋白及其重链的研制、特性鉴定和应用

目的 分离、提纯人心肌肌球蛋白(HCM)及其重链(HCMHC),为心肌显像及其显像剂抗HCMHC抗体的研究打基础。方法 参考猪心肌肌球蛋白制备方法,做了较大改进后,提取了HCM,用盐酸胍裂解HCM制备了HCMHC。用SDS-PAGE(梯度)电泳测定分子质量,用检测无机磷的方法测定HCM的ATP酶活力,用Ellman氏试剂测定游离巯基。将HCMHC免疫动物,用ELISA法测定了血清抗体效价。结果 HCMHC的相对分子量为200×10³,纯度大于80%;被免疫的BALB/C小鼠血清的抗体滴度最高达10⁷,细胞融合率达80%。结论 研制的HCMHC具有良好的免疫学活性,为下一步抗HCMHC单链可变区抗体(ScFv)及其放射性核素受体显像的研究打下了基础。

Studies on Preparation, Characteristic Appraisal and Application of Human Cardiac Myosin and Human Cardiac Myosin Heavy Chain

Objective To isolate,extract and purify human cardiac myosin(HCM)and human cardiac myosin heavy chain(HCMHC)for it's next study of myocardid image and the antibody against HCMHC imaging reagent.Methods First extract HCM from human heart muscle tissue and prepare HCMHC from the HCM,detecte characteristics including protein concentration,molecular weight,enzymatic activity and free SH groups of the HCM and the HCMHC,prepare hybridoma cell lines of HCMHC,and examine the immunology activity of the HCMHC.Results 24mg HCM were extracted from 127g human cardiac muscle tissue,and 9.1mg HCMHC were prepared.Elecrtrophoresis of SDS-PAGE(gradient)showed that the band of HCMHC located at 200 kD,which was the same of that of literatures,purity of HCMHC is higher than 80%.The specific activity of ATPase of HCM is good.The highest titer of anti-HCMHC serum is about 107.The fusion rate was 80%.Conclusions HCMHC has good immunology activity,for it's next study of the antibody against HCMHC single chain fragment variable(ScFv)radionuclide receptor imaging.