DNA aptamer-based affinity chromatography system for purification of recombinant proteins tagged with lysine tag.

Autor: Arciszewska K; Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland., Kowalska E; Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland., Bartnicki F; Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland., Bonarek P; Department of Physical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland., Banaś AK; Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland., Strzałka W; Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland. Electronic address: wojciech.strzalka@uj.edu.pl.
Jazyk: angličtina
Zdroj: Journal of chromatography. A [J Chromatogr A] 2023 Mar 15; Vol. 1692, pp. 463846. Date of Electronic Publication: 2023 Feb 07.
DOI: 10.1016/j.chroma.2023.463846
Abstrakt: Affinity chromatography (AC) is one of the techniques widely used for the purification of recombinant proteins. In our previous study, we presented a successful application of the Argi system [1] for the purification of recombinant proteins, based on the specific interaction between an arginine tag and a DNA aptamer. Exploring the possible application of positively charged peptide tags in the purification of recombinant proteins, in this study we developed and characterized an AC system based on the specific and reversible interaction between a DNA aptamer and a lysine tag (Lys-tag) comprising five lysine residues (5 K). We optimized the length of both the selected DNA aptamer and Lys-tag which were named B5K aptamer and 5K-tag, respectively. The results showed that the stability of the B5K aptamer and 5K-tag was dependent on the presence of potassium ions. The conditions for mild elution of 5K-tagged protein from B5K aptamer were determined. Our study proved that the developed system can be used for the purification of recombinant proteins from Escherichia coli total protein extracts.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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