DNA aptamer-based affinity chromatography system for purification of recombinant proteins tagged with lysine tag. 2023

Klaudia Arciszewska, and Ewa Kowalska, and Filip Bartnicki, and Piotr Bonarek, and Agnieszka Katarzyna Banaś, and Wojciech Strzałka
Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, Krakow 30-387, Poland.

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.

UI MeSH Term Description Entries
D007202 Indicators and Reagents Substances used for the detection, identification, analysis, etc. of chemical, biological, or pathologic processes or conditions. Indicators are substances that change in physical appearance, e.g., color, at or approaching the endpoint of a chemical titration, e.g., on the passage between acidity and alkalinity. Reagents are substances used for the detection or determination of another substance by chemical or microscopical means, especially analysis. Types of reagents are precipitants, solvents, oxidizers, reducers, fluxes, and colorimetric reagents. (From Grant & Hackh's Chemical Dictionary, 5th ed, p301, p499) Indicator,Reagent,Reagents,Indicators,Reagents and Indicators
D008239 Lysine An essential amino acid. It is often added to animal feed. Enisyl,L-Lysine,Lysine Acetate,Lysine Hydrochloride,Acetate, Lysine,L Lysine
D011993 Recombinant Fusion Proteins Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes. Fusion Proteins, Recombinant,Recombinant Chimeric Protein,Recombinant Fusion Protein,Recombinant Hybrid Protein,Chimeric Proteins, Recombinant,Hybrid Proteins, Recombinant,Recombinant Chimeric Proteins,Recombinant Hybrid Proteins,Chimeric Protein, Recombinant,Fusion Protein, Recombinant,Hybrid Protein, Recombinant,Protein, Recombinant Chimeric,Protein, Recombinant Fusion,Protein, Recombinant Hybrid,Proteins, Recombinant Chimeric,Proteins, Recombinant Fusion,Proteins, Recombinant Hybrid
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002846 Chromatography, Affinity A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed) Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D052157 Aptamers, Nucleotide Nucleotide sequences, generated by iterative rounds of SELEX APTAMER TECHNIQUE, that bind to a target molecule specifically and with high affinity. DNA Aptamer,DNA Aptamers,RNA Aptamers,Rna Aptamer,Nucleotide Aptamers,Oligonucleotide Ligands, DNA,Oligonucleotide Ligands, RNA,Aptamer, DNA,Aptamer, Rna,Aptamers, DNA,Aptamers, RNA,DNA Oligonucleotide Ligands,RNA Oligonucleotide Ligands

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