人心肌肌钙蛋白I的纯化及鉴定

Purification and identification of cardiac troponin I from human myocardium

目的 提取和纯化人心肌肌钙蛋白I。方法 心室肌经匀浆、离心、盐析提取 ,CMSephadexC 5 0柱层析 ,DEAESephadexA 5 0柱层析制备人cTnI ;经 (cTnI)活性检测、SDS PAGE、westernblot鉴定。结果 从 6 5 g湿重心肌中获得 4 .1mgcTnI ,分子量约为 2 4 0 0 0 ,纯度约为 84 %。结论 纯化的cTnI可被cTnI单克隆抗体特异识别 ,本实验从人心肌中成功制备cTnI。

Objective To extract and purify cardiac troponin I (cTnI) from human myocardium.Methods The cTnI was separated from human myocardium by homogenization,centrifugation, salting-out and precipitation by ammonium sulfate, and CM-Sephadex C-50 and DEAE Sephadex A-50 column chromatography.The purified cTnI was identified by SDS-PAGE and Western Blot.Results The total yield of cTnI obtained from 65g human cardiac muscle was 4.1mg. Its molecular mass was approximately 24kDa and purity was 84%. The purified cTnI showed specific binding affinity to anti-cTnI monoclonal antibody.Conclusion The present method can be used as an approach to prepare purified cTnI from cardiac muscle.