title: Aptamer-based downstream processing of his-tagged proteins utilizing magnetic beads.
abstract: Aptamers are synthetic nucleic acid-based high affinity ligands that are able to capture their corresponding target via molecular recognition. Here, aptamer-based affinity purification for His-tagged proteins was developed. Two different aptamers directed against the His-tag were immobilized on magnetic beads covalently. The resulting aptamer-modified magnetic beads were characterized and successfully applied for purification of different His-tagged proteins from complex E. coli cell lysates. Purification effects comparable to conventional immobilized metal affinity chromatography were achieved in one single purification step. Moreover, we have investigated the possibility to regenerate and reuse the aptamer-modified magnetic beads and have shown their long-term stability over a period of 6 months.
authors.affiliation: Institut für Technische Chemie, Leibniz Universität Hannover, Callinstrasse 5, 30167 Hannover, Germany; telephone: +49-511-762-2968; fax: +49-511-762-3004.
authors.forename: Öznur
authors.initials: Ö
authors.lastname: Kökpinar
authors.forename: Johanna-Gabriela
authors.initials: JG
authors.lastname: Walter
authors.forename: Yuval
authors.initials: Y
authors.lastname: Shoham
authors.forename: Frank
authors.initials: F
authors.lastname: Stahl
authors.forename: Thomas
authors.initials: T
authors.lastname: Scheper
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D006639:Histidine / Q000737:chemistry*; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D052157:Aptamers, Nucleotide; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins; C033223:polyhistidine; D006639:Histidine