title: Nickel(II)-immobilized sulfhydryl cotton fiber for selective binding and rapid separation of histidine-tagged proteins.
abstract: In the current study, a novel nickel(II)-immobilized sulfhydryl cotton fiber (SCF-Ni(2+)) was prepared in a simple way based on the coordination effect between Ni(2+) and thiol group on the surface of SCF. The composition and element mapping of SCF-Ni(2+) fibers were demonstrated by energy-dispersive X-ray (EDX) spectroscopy. Based on the high affinity of Ni(2+) to 6×His on histidine-tagged (His-tagged) proteins, SCF-Ni(2+) fibers were then further used as an immobilized metal ion affinity chromatography (IMAC) adsorbent for selective binding and rapid separation of His-tagged proteins using an in- pipette-tip SPE format. Our results showed that SCF-Ni(2+) adsorbent can selectively capture His-tagged proteins from protein mixture and Escherichia coli cell lysates. Taken together, the developed method provides a rapid, convenient and efficient approach for the purification of His-tagged proteins.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, P.R. China.
authors.forename: Xiao-Mei
authors.initials: XM
authors.lastname: He
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, P.R. China.
authors.forename: Gang-Tian
authors.initials: GT
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Lu
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, P.R. China.
authors.forename: Bi-Feng
authors.initials: BF
authors.lastname: Yuan
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, P.R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan 430072, P.R. China. Electronic address: yqfeng@whu.edu.cn.
authors.forename: Yu-Qi
authors.initials: YQ
authors.lastname: Feng
mesh_terms: D002846:Chromatography, Affinity; D056831:Coordination Complexes / Q000737:chemistry*; D032561:Cotton Fiber; D004926:Escherichia coli / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006639:Histidine / Q000737:chemistry*; D009532:Nickel / Q000737:chemistry*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D013438:Sulfhydryl Compounds
chemical_list: D056831:Coordination Complexes; D011994:Recombinant Proteins; D013438:Sulfhydryl Compounds; D049452:Green Fluorescent Proteins; D006639:Histidine; D009532:Nickel
keywords: E. coli cell lysates; His-tagged proteins; Immobilized metal ion affinity chromatography (IMAC); Ni(2+)-immobilized sulfhydryl cotton fiber (SCF-Ni(2+))