IMBICE   05372
INSTITUTO MULTIDISCIPLINARIO DE BIOLOGIA CELULAR
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Use of palladium ions for recombinant proteins recovery
Autor/es:
PAMELA KIKOT; MARCELO FERNANDEZ LAHORE; MARIANO GRASSELLI
Lugar:
Chascomús
Reunión:
Encuentro; IV Latin American Meeting on Biological Inorganic Chemistry; 2014
Institución organizadora:
UBA
Resumen:
Peptide-based tags fusion to recombinant proteins is nowadays one of the most used tools developed for protein production in research labs. The combination of expression systems of recombinant tagged proteins with the immobilized metal ion affinity chromatography (IMAC) became in an efficient and rapid tool to produce milligram-range amounts of proteins1. IMAC-Ni(II) column is the natural partner of the 6xHis tagged proteins, but also other metal ions have been used with other amino acid specificities. The Pd(II) ion is able to form ternary complexes with iminodiacetic acid (IDA) and side chains of free amino acids2;3. However, according our acknowledge, the successful use of IMAC-Pd(II) to purify tagged proteins has not been reported yet. In this work is described the application of Pd(II) ion to IMAC as an alternative purification strategy. We evaluated different Cys- and His-containing six-amino-acid tags linked to the N-terminal group of GFP, as a model protein,and studied the adsorption and elution conditions using different reagents. Cys-containing tagged GFPs were able to bind to IMAC-Pd(II) matrices and eluted successfully using a thiourea solution. We also determined the purification performance of a Cys-containing tagged GFP from a crude homogenate of E. coli.