title: Establishment of a three-step purification scheme for a recombinant protein rG17PE38 and its characteristics identification.
abstract: A protein with high purity has become an essential pre-requisite for investigating its bioactivity, molecular structure and characteristics. Therefore, the development of technologies for efficient purification of protein is urgently necessary. The objective of this study was to establish a purification protocol for a recombinant protein rG17PE38. Different forms of chromatography such as hydrophobic interaction and ion exchange chromatography were chosen as the core purification steps. The performance of each technique was optimized to meet the requirements and the purification steps were arranged in a logical way of facilitating to operate in next step. In addition, some characteristics of the protein such as stability, bioactivity and cellular location were determined. Finally, whether the protein could induce cell apoptosis was also explored. The results showed the protein purified via the suggested three-step purification scheme could obtain a purity of 95%, and its bioactivity in the form of IC₅₀ was 17.6 ng/mL, furthermore it could keep stable at 4 °C for at least 10 days. The protein could bind on its target cell membrane specifically, and inducing cell apoptosis was demonstrated to be one of the cytotoxicity mechanisms of the protein. Results obtained in our study may provide useful information on strategies of protein purification and lay a substantial foundation for the followed animal or clinical experiments on rG17PE38.
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Feng
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Zeng-Shan
authors.initials: ZS
authors.lastname: Liu
authors.affiliation: China-Japan Union Hospital, Jilin University, Changchun 130033, China.
authors.forename: Xi-Lin
authors.initials: XL
authors.lastname: Liu
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Shi-Ying
authors.initials: SY
authors.lastname: Lu
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Yan-Song
authors.initials: YS
authors.lastname: Li
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Pan
authors.initials: P
authors.lastname: Hu
authors.affiliation: Wuhu Kang Wei Biological Technology Co., Ltd., Wuhu 241000, China.
authors.forename: Dong-Ming
authors.initials: DM
authors.lastname: Yan
authors.affiliation: Department of General Surgery, The First Hospital, Jilin University, Changchun 130021, China.
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Tong
authors.affiliation: Department of General Surgery, The First Hospital, Jilin University, Changchun 130021, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Jin
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Yan-Xia
authors.initials: YX
authors.lastname: Ding
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Dong-Xue
authors.initials: DX
authors.lastname: Gai
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China. Electronic address: renhl@jlu.edu.cn.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Ren
mesh_terms: D000645:Ammonium Sulfate; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D011232:Chemical Precipitation; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; D000645:Ammonium Sulfate
keywords: Apoptosis; Chromatography; Purification; Recombinant protein