文献类型：期刊文献

中文题名：基于铁蛋白的溶栓蛋白纳米粒子的构建及活性分析

英文题名：Construction and Activity Analysis of Ferritin-Based Thrombolytic Protein Nanoparticles

作者：赵剑[1];沈阳[1];曹旭妮[1]

机构：[1]华东理工大学生物工程学院生物反应器工程国家重点实验室

年份：2019

卷号：45

期号：4

起止页码：576

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;Scopus;北大核心:【北大核心2017】；CSCD:【CSCD_E2019_2020】；

基金：国家自然科学基金面上项目(21375039)

语种：中文

中文关键词：单链尿激酶型纤溶酶原激活剂;铁蛋白重链亚基;蛋白纳米粒子;溶栓

外文关键词：single chain urokinase-type plasminogen activator;ferritin H-chain protein;protein nanoparticles;thrombolysis

摘要：将凝血酶激活的低分子量单链尿激酶型纤溶酶原激活剂(T-uPA)修饰铁蛋白重链亚基(FTH1),构建基于铁蛋白的溶栓蛋白纳米粒子.通过基因工程、体外混合复性等技术,将T-uPA分别修饰FTH1的N和C端并进行对比,表明仅C端融合蛋白才可复性折叠并组装成FTH1/FTH1-T-uPA.通过实验分析,表明FTH1/FTH1-T-uPA基本为中空笼状且分散均匀,粒径为13.41 nm,水合粒径为24.2 nm,T-uPA可成功被凝血酶激活,具有优良的纤维蛋白溶解活性(约1200 IU/mg);在体外达到与商品化尿激酶相似的溶栓效果,且具有更好的安全性.本研究成功解决了T-uPA修饰铁蛋白的关键问题,为构建靶向型的铁蛋白纳米粒子载药系统提供了基础.
Thrombotic diseases have led to seriously threat to human health. In recent years, the development goal of thrombolytic drugs have obtained new thrombolytic agents with good thrombolytic effect and highly drug safety. Upon using ferritin as the carrier, thrombolytic protein nanoparticles were constructed by modifying human ferritin Hchain protein (FTH1) with the low molecular weight single chain urokinase-type plasminogen activator which can be activated by thrombin (T-uPA). T-uPA was modified at the N terminus and C terminus of FTH1, respectively, using genetic engineering, in vitro mixed renaturation technology and so on. It was found that only the T-uPA modified at the FTH1 (FTH1-T-uPA) could be folded correctly and assembled into the FTH1/FTH1-T-uPA. A series of characterizations showed that the FTH1 and FTH1-T-uPA subunits were successfully folded and assembled into the characteristic cages of ferritin. The FTH1/FTH1-T-uPA was obtained in the shape of spherical particles with a diameter of 13.41 nm and presented a very narrow size distribution in solution with a hydrodynamic diameter of 24.2 nm. Importantly, the T-uPA molecules of FTH1-T-uPA could be successfully activated by thrombin and had excellent fibrinolytic activity (1 200 IU/mg). The FTH1/FTH1-T-uPA could achieve comparable thrombolytic effects with the commercial urokinase, and showed much better safety in vitro. This study shows that T-uPA can be successfully modified on the surface of ferritin and provide the methods for designing targeted drug loading system on the basis of ferritin nanoparticles.