摘要
目的以大肠埃希菌BL21表达的重组人胱抑素C(CysC)蛋白作为原材料制备CysC质控品,并对制备的质控品的各项性能进行分析。方法通过基因工程手段,利用表达载体p Cold1及宿主大肠埃希菌BL21表达重组人CysC蛋白,纯化后对其进行冷冻干燥。结果成功表达出重组人CysC蛋白包涵体,透析复性后得到具有免疫学活性的可溶性CysC重组蛋白。利用原核表达的CysC蛋白制备CysC质控品,并经过HITACHI及Roche平台进行检测,CysC质控品在这2个平台都适用。结论利用重组人CysC蛋白制备的CysC质控品具有良好的稳定性及可靠性,可以作为质控品用于不同的体外检测平台。
Objective To prepare cystatin C(Cys C)quality control material by using recombinant human Cys C protein expressed in Escherichia coli BL21as basic material.Methods By gene engineering method,recombinant human Cys C protein was expressed through pCold1and BL21.Freeze drying was performed after purification.Results Inclusion bodies of recombinant human Cys C protein was expressed successfully.After renaturation by dialysis method,soluble recombinant human Cys C protein with immunological activity had been prepared.Cys C quality control material was prepared by using the recombinant human Cys C protein and determined by HITACHI and ROCHE analysis systems.Cys C quality control was suitable for the2analysis systems.Conclusions Cys C quality control material prepared by recombinant human Cys C protein has good stability and reliability,it could be used in various analysis platforms of in vitro diagnosis.
作者
李江峰
冯敏仪
钱伟
赵晓妮
陈梓彤
矫丽媛
王继华
才蕾
LI Jiangfeng;FENG Minyi;QIAN Wei;ZHAO Xiaoni;CHEN Zitong;JIAO Liyuan;WANG Jihua;CAI Lei(National and Local United Engineering Laboratory of High-Efficiency Self-Testing Reagents,Quality Control Laboratory of Research and Development Center of Guangzhou Wondfo Biotech Co.,Ltd.,Guangzhou 510641,Guangdong,China)
出处
《检验医学》
CAS
2017年第8期733-737,共5页
Laboratory Medicine
关键词
胱抑素C
重组蛋白
质控品
稳定性
Cystatin C
Recombinant protein
Quality control material
Stability
