title: Simple enrichment of thiol-containing biomolecules by using zinc(II)-cyclen-functionalized magnetic beads.
abstract: A simple and efficient method based on magnetic-bead technology has been developed for the enrichment of thiol-containing biomolecules, such as l-glutathione and cysteine-containing peptides. The thiol-binding site on the bead is a mononuclear complex of zinc(II) with 1,4,7,10-tetraazacyclododecane (cyclen); this is linked to a hydrophilic cross-linked agarose coating on a particle that has a magnetic core. All steps for the thiol-affinity separation are conducted in aqueous buffers with 0.10 mL of the magnetic beads in a 1.5 mL microtube. The entire separation protocol for thiol-containing compounds, from addition to elution, requires less than one hour per sample, provided the buffers and the zinc(II)-cyclen-functionalized magnetic beads have been prepared in advance. The thiol-affinity magnetic beads are reusable at least 15 times without a decrease in their thiol-binding ability, and they are stable for six months at room temperature.
authors.affiliation: Laboratory of Organic Medicinal Chemistry, Faculty of Pharmacy & Pharmaceutical Sciences, Fukuyama University, Fukuyama, Japan.
authors.forename: Haruto
authors.initials: H
authors.lastname: Fujioka
authors.forename: Masaya
authors.initials: M
authors.lastname: Tsunehiro
authors.forename: Maho
authors.initials: M
authors.lastname: Kawaguchi
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kuramoto
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Kurosaki
authors.forename: Yuhzo
authors.initials: Y
authors.lastname: Hieda
authors.forename: Emiko
authors.initials: E
authors.lastname: Kinoshita-Kikuta
authors.forename: Eiji
authors.initials: E
authors.lastname: Kinoshita
authors.forename: Tohru
authors.initials: T
authors.lastname: Koike
mesh_terms: D000080027:Cyclams; D005978:Glutathione / Q000737:chemistry / Q000302:isolation & purification*; D006571:Heterocyclic Compounds / Q000737:chemistry*; D060328:Magnetic Phenomena; D008280:Magnetics / Q000295:instrumentation / Q000379:methods*; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D015032:Zinc / Q000737:chemistry*
chemical_list: D000080027:Cyclams; D006571:Heterocyclic Compounds; D010455:Peptides; C038072:cyclen; D005978:Glutathione; D015032:Zinc
keywords: Affinity enrichment; Cysteine-containing peptide; Magnetic beads; Thiols; Zinc