聚乙二醇单修饰的重组人粒细胞集落刺激因子突变体的纯化与分析  

Purification and analysis of rmhG-CSF mutant modified with polyethylene glycol

作　　者：张金玲[1,2] 马南佳[1] 汪军远[1] 方井晋[1] 刘晓妮[1] 

机构地区：[1]杭州九源基因工程有限公司,浙江杭州310018 [2]浙江工业大学,浙江杭州310024

出　　处：《华西药学杂志》2010年第2期129-131,共3页West China Journal of Pharmaceutical Sciences

摘　　要：目的获得高活性的聚乙二醇定点单修饰的产物PEG-rmhG-CSF-Cys176。方法用聚乙二醇马来酰亚胺基和高活性的重组人粒细胞集落刺激因子突变体(rmhG-CSF)的半胱氨酸(Cys)基团特异性结合,并用离子交换层析分离纯化修饰产物,通过高效凝胶过滤色谱检测纯度,MTT法检测体外的生物活性。结果蛋白纯度为97%,体外生物活性为0.8×108IU·mg-1。结论半胱氨酸反应性PEG和游离Cys的结合是在rmhG-CSF活性区以外的位置C末端,修饰后的蛋白活性基本未受影响。OBJECTIVE To obtain a high active polyethylene glycol-modified product of fixed-point single-PEG-rmhG-CSF-Cys176.METHODS PEG-based maleimide (mPEG-Maleimide,mPEG-Mal)was conjugated with highly active recombinant human granulocyte colony-stimulating factor mutant (rmhG-CSF) cysteine (Cys) group.The products were separated and purified by ion-exchange chromatography.The purity of product was determined by high performance gel filtration chromatography and in vitro biological activity was determined by MTT met...

关 键 词：重组人粒细胞集落刺激因子 突变体 聚乙二醇 高效凝胶过滤色谱 

分 类 号：R55[医药卫生—血液循环系统疾病]