IMMOBILIZED PALLADIUM ION AFFINITY CHROMATOGRAPHY FOR RECOVERY HIS-TAG PROTEINS

KIKOT, PAMELA; MARCELO FERNANDEZ LAHORE; MARIANO GRASSELLI

Rosario

Congreso; CAIQ2013 - VII CONGRESO ARGENTINO DE INGENIERíA QUíMICA; 2013

Asociacion Argentina de Ingenieros Quimicos

Abstract. Develop novel tools and solutions for protein purification are always welcome in the scientific or research laboratories. One of the most successful concepts developed in the field of affinity chromatography was its combination with peptide-based tags. Immobilized metal ion affinity chromatography (IMAC) is used to purify proteins with a His tag added at its N o C terminal sequence (an amino acid with the strongest affinity for metal ions). The combination of expression systems of recombinant tagged proteins with a robust chromatographic system became in an efficient and rapid tool to produce milligram-range amount of protein. Commercial protein-compatible chromatographic matrices based on Pd2+ interaction have not been studied as protein purification tool of recombinant His-tag proteins. In this report it is compared the adsorption/desorption properties of a His-tagged protein, green fluorescent protein, in IMAC-Pd2+ and IMACNi2+ system. Selection of eluent, chromatographic behavior and protein recovery yield are also analyzed.