文献类型：期刊文献

英文题名：Soluble Expression of Recombinant Human Cystatin C and Comparison of the Ni Column and Magnetic Bead Purification

作者：Zhang, Yibin[1];Zhao, Jian[1];He, Shiyu[1];Cao, Xuni[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2020

卷号：39

期号：1

起止页码：85

外文期刊名：PROTEIN JOURNAL

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000490839200001)】；

基金：This study was supported by the National Natural Science Foundation of China (Grant No. 21375039), the Shanghai Committee of Science and Technology (Grant Nos. 16142202000 and 18142202600), the Research Program of Shanghai Chemical Reagent Research Institute Co., Ltd. (Grant F100-81707), and the National Special Fund for State Key Laboratory of Bioreactor Engineering (Grant No. 2060204).

语种：英文

外文关键词：Cystatin C; Efficient purification; Magnetic beads; Periplasmic expression

摘要：Cystatin C, also known as gamma-trace or post-gamma-globulin, is a cysteine protease inhibitor from the cystatin superfamily. It is usually used as a marker of the glomerular filtration rate owing to its low molecular weight and constant secretion. The recently available methods for cystatin C preparation have low outputs. Hence, a productive preparation system is urgently required. In this study, a 6 x His-tag coupled with a thrombin cleavage site was fused to the C-terminus of cystatin C, and the protein was well expressed in Escherichia coli after optimization. Then, two different systems were used to obtain no-tag cystatin C: a traditional nickel (Ni)-column system and a subtly Ni magnetic bead system. The column system was more commonly used, and the magnetic bead system was more convenient. Cystatin C (purity > 97%) was successfully obtained, and the yields in both the systems were higher than those in previous studies. Further, the proper folding status and bioactivity of recombinant cystatin C were confirmed using the papain inhibition assay, dynamic light scattering, and circular dichroism spectroscopy.