title: [Isolation and purification of enhanced green fluorescent protein using chromatography].
abstract: Enhanced green fluorescent protein (EGFP) is a common biological marker. In this research, on the foundation of successful clone and expression of EGFP, a two-step chromatographic method was established to separate and purify EGFP, which includes the use of HisTrap HP immobilized metal affinity chromatography (IMAC) and Sephadex G-10 HR size exclusion chromatography in sequence. Sephacryl S-300 HR size exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to check out the purity of EGFP. At last, it was found that EGFP still had fluorescent activity using fluorescence spectrophotometric detection and Native-PAGE detection. This method can effectively separate the active EGFP. The purity of the obtained EGFP was more than 98%.
authors.affiliation: Weihai International Biotechnology R&D Center, Shandong University, Weihai 264209, China.
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Hou
authors.forename: Shuliang
authors.initials: S
authors.lastname: Song
authors.forename: Hao
authors.initials: H
authors.lastname: Liang
authors.forename: Weili
authors.initials: W
authors.lastname: Wang
authors.forename: Aiguo
authors.initials: A
authors.lastname: Ji
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D005456:Fluorescent Dyes / Q000302:isolation & purification; D049452:Green Fluorescent Proteins / Q000302:isolation & purification*
chemical_list: D005456:Fluorescent Dyes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins