title: Imidazole-free purification of His3-tagged recombinant proteins using ssDNA aptamer-based affinity chromatography.
abstract: Immobilized metal ion affinity chromatography (IMAC) is widely used for the purification of many different His6-tagged recombinant proteins. On the one hand, it is a powerful technique but on the other hand it has its disadvantages. In this report, we present the development of a unique ssDNA aptamer for the purification of His3-tagged recombinant proteins. Our study shows that stability of the His3-tag/H3T aptamer complex can be controlled by the sodium ion concentration. Based on this feature, we demonstrate that H3T aptamer resin was successfully employed for the purification of three out of four tested His3-tagged recombinant proteins from an E. coli total protein extract using imidazole-free buffers. Finally, we show that the purity of His3-tagged proteins is superior when purified with the help of the H3T aptamer in comparison with Ni-NTA resin.
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors.forename: Filip
authors.initials: F
authors.lastname: Bartnicki
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Kowalska
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Pels
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland. Electronic address: wojciech.strzalka@uj.edu.pl.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Strzalka
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry*; D001483:Base Sequence; D002846:Chromatography, Affinity / Q000379:methods; D004277:DNA, Single-Stranded / Q000737:chemistry*; D004926:Escherichia coli / Q000235:genetics; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D007202:Indicators and Reagents; D011994:Recombinant Proteins; D006639:Histidine
keywords: DNA aptamers; Histidine tag; IMAC; Imidazole; SELEX