an automatic refolding apparatus for preparative-scale protein production自动重折叠蛋白质制备规模生产装置.pdfVIP

An Automatic Refolding Apparatus for Preparative-Scale Protein Production 1,2 1 2 1 3 1 Yanye Feng , Ming Zhang , Linlin Zhang , Ting Zhang , Jianfeng Ding , Yingping Zhuang , 1 2 Xiaoning Wang *, Zhong Yang * 1 State Key Laboratory of Bioreactor Engineering, School of Biotechnology, East China University of Science and Technology, Shanghai, China, 2 State Key Laboratory of Genetic Engineering, Department of Microbiology, School of Life Sciences, Fudan University, Shanghai, China, 3 Department of RD, Novoprotein Scientific Inc, Shanghai, China Abstract Protein refolding is an important process to recover active recombinant proteins from inclusion bodies. Refolding by simple dilution, dialysis and on-column refolding methods are the most common techniques reported in the literature. However, the refolding process is time-consuming and laborious due to the variability of the behavior of each protein and requires a great deal of trial-and-error to achieve success. Hence, there is a need for automation to make the whole process as convenient as possible. In this study, we invented an automatic apparatus that integrated three refolding techniques: varying dilution, dialysis and on-column refolding. We demonstrated the effectiveness of this technology by varying the flow rates of the dilution buffer into the denatured protein and testing different refolding methods. We carried out different refolding methods on this apparatus: a combination of dilution and dialysis for human stromal cell-derived factor 1 (SDF-1/ CXCL12) and thioredoxin fused-human artemin protein (Trx-ARTN); dilution refolding for thioredoxin fused-human insulin- like growth factor I protein (Trx-IGF1) a