论文部分内容阅读
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。
Objective To isolate and purify human cardiac myosin (HCM) and its heavy chain (HCMHC) and lay the foundation for the study of myocardial imaging and its imaging agent against HCMHC. Methods With reference to preparation method of porcine myocardium myosin, HCMHC was extracted after HCMHC was extracted and HCMHC was cleaved with guanidine hydrochloride. The molecular mass was determined by SDS-PAGE (gradient) electrophoresis, the ATPase activity of HCM was measured by the method of detecting inorganic phosphorus, and the free thiol group was measured by Ellman’s reagent. The animals were immunized with HCMHC and serum antibody titers were determined by ELISA. Results The relative molecular weight of HCMHC was 200 × 103 and the purity was more than 80%. The antibody titer of the immunized BALB / C mice was up to 107 and the cell fusion rate was 80%. Conclusion The developed HCMHC has good immunological activity, which lays the foundation for the further study on the scFv antibody and its radionuclide receptor imaging.