title: Evaluation of immobilized metal affinity chromatography kits for the purification of histidine-tagged recombinant CagA protein.
abstract: Immobilized metal affinity chromatography (IMAC) technique is used for fast and reliable purification of histidine(His)-tagged recombinant proteins. The technique provides purification under native and denaturing conditions. The aim of this study is to evaluate three commercially available IMAC kits (Thermo Scientific, GE Healthcare and Qiagen) for the purification of a 6xHis-tagged recombinant CagA (cytotoxin-associated gene A) protein from IPTG-induced Escherichia coli BL21(DE3) culture. The kits were tested according to the manufacturer instructions and the protein was purified with only GE Healthcare and Qiagen kits under denaturing conditions. 1% (w/v) SDS was used as denaturing agent in PBS instead of extraction reagent of Thermo Scientific kit to lyse bacterial cells from 100ml culture. The 6xHis-tagged recombinant protein was purified by the three kits equally.
authors.affiliation: Fatih University, Faculty of Science and Arts, Department of Biology, 34500 Buyukcekmece, Istanbul, Turkey. Electronic address: cebrail4u@gmail.com.
authors.forename: Cebrail
authors.initials: C
authors.lastname: Karakus
authors.affiliation: Fatih University, Faculty of Science and Arts, Department of Biology, 34500 Buyukcekmece, Istanbul, Turkey.
authors.forename: Merve
authors.initials: M
authors.lastname: Uslu
authors.affiliation: Fatih University, Faculty of Science and Arts, Department of Biology, 34500 Buyukcekmece, Istanbul, Turkey.
authors.forename: Duygu
authors.initials: D
authors.lastname: Yazici
authors.affiliation: Fatih University, Faculty of Science and Arts, Department of Biology, 34500 Buyukcekmece, Istanbul, Turkey.
authors.forename: Barik A
authors.initials: BA
authors.lastname: Salih
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D006639:Histidine / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011994:Recombinant Proteins; C080866:cagA protein, Helicobacter pylori; D006639:Histidine
keywords: 6xHis-tag; Denaturing conditions; IMAC; Immobilized metal affinity chromatography; Protein purification; Recombinant protein; SDS