title: Expression and purification of bioactive high-purity recombinant mouse SPP1 in Escherichia coli.
abstract: Secreted phosphoprotein 1 (SPP1) is a phosphorylated acidic glycoprotein. It is broadly expressed in a variety of tissues, and it is involved in a number of physiological and pathological events, including cancer metastasis, tissues remodeling, pro-inflammation regulation, and cell survival. SPP1 has shown its function of protecting tissues and organs against injury and wound, giving itself potentials to become a therapy target or giving its antibodies of other counter-acting reagents potentials to become drug candidates. Non-tagged (native) recombinant SPP1 would be valuable in therapeutic and pharmaceutical researches. In our study, mouse Spp1 DNA fragment without signal peptide was built in pET28a(+) vector and transformed into Escherichia coli BL21 (DE3). The recombinant mouse SPP1 (rmSPP1) was then expressed in bacteria upon induction by isopropyl β-D-thiogalactopyranoside (IPTG). The abundance of rmSPP1 was increased using isoelectric precipitation and ammonium sulfate fractionation methods, and anion and cation exchange chromatography was employed to further purify rmSPP1. Finally, we got rmSPP1 product with 12.8 % productivity, 97 % purity, satisfactory bioactivity, and low endotoxin content.
authors.affiliation: Shanghai Key Laboratory of Veterinary Biotechnology, College of Agriculture and Biology, Shanghai Jiao Tong University, 800 Dongchuan Rd, 200240, Shanghai, People's Republic of China, yunsheng@sjtu.edu.cn.
authors.forename: Yunsheng
authors.initials: Y
authors.lastname: Yuan
authors.forename: Xiyuan
authors.initials: X
authors.lastname: Zhang
authors.forename: Shunyan
authors.initials: S
authors.lastname: Weng
authors.forename: Wen
authors.initials: W
authors.lastname: Guan
authors.forename: Di
authors.initials: D
authors.lastname: Xiang
authors.forename: Jin
authors.initials: J
authors.lastname: Gao
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Han
authors.forename: Yan
authors.initials: Y
authors.lastname: Yu
mesh_terms: D000818:Animals; D002633:Chemotaxis / Q000187:drug effects; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D008099:Liver / Q000201:enzymology; D051379:Mice; D053495:Osteopontin / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; C506896:Spp1 protein, mouse; D053495:Osteopontin