摘要
为探讨钴金属螯合亲和层析应用于分离纯化制备人胰岛素样生长因子-1(hIGF-1)的可行性与纯化条件,以重组工程菌E.coli DH5α/pET32a(IGF-1)为对象,使用钴亲和层析技术分离纯化其表达产物并与镍亲和层析比较,在此基础上尝试应用钴亲和层析分离纯化制备较大量的hIGF-1融合蛋白。结果显示,钴亲和层析在分离纯化hIGF-1融合蛋白时表现出更好的可操作性与纯化效果,线性放大条件后分离纯化效果稳定,制备hIGF-1融合蛋白产物纯度约为95%,蛋白获得率约为10.78%。研究初步建立了切实可行的钴金属螯合亲和层析分离纯化制备hIGF-1融合蛋白的工艺。
To study the feasibility and appropriate conditions for the purification of recombinant human insulin-like growth factor-1 （ hIGF-1 ）, Metal Chelate Affinity Chromatography （ MCAC ） with Cobalt ion （ Co2＋） compared with Nickel ion （ Ni2 ＋ ） was carried out for purifying hIGF-1 fusion protein expressed in E. coli DI-ISo./pET32a （ IGF-1 ）. Then Co-purification was used to make the large-scale preparation of purified hIGF-1 fusion protein by linear amplification. The results showed that Co-purification exhibited the better maneuverability and higher selectivity of the target protein, and its effect was stable after linear amplification. By analysis of SDS-PAGE electrophoresis gel it could be demonstrated that the purity of hIGF-1 fusion protein was about 95%, and the obtained rate was about 10.78%. The study established a practical method for the protein production process of hlGF-1 fusion protein purified by MCAC with Co2＋.
出处
《药物生物技术》
CAS
2013年第3期220-224,共5页
Pharmaceutical Biotechnology
基金
福建省高校产学合作科技重大项目(No.2010Y4002)
