CAMPERI Silvia Andrea
congresos y reuniones científicas
Bio-layer interferometry using optic-based biosensor for screening of affinity ligand for rhEPO.
M. C. MARTÍNEZ-CERON; M. M. MARANI; M. ETCHEVERRIGARAY; M. TAULÉS; F. ALBERICIO; O. CASCONE; S. A. CAMPERI
Congreso; XLVI Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Bio-layer interferometry is used in optic-based biosensors (ForteBio OCTET RED) for protein-ligands interactions analysis. Short peptides are ideal to be used as affinity ligands for protein purification. Divide-couple-recombine (DCR) method allows obtaining a library with all possible combinations of the amino acids in the form of "one bead-one peptide". An octapeptide library XXXOXXOO (O=fixed positions and X =Ala, Asp, Glu, Phe, His, Leu, Asn, Pro, Ser y Thr) was synthesised. After the library screening 60 peptides with affintiy for rhEPO were selected. To study the peptide affinity for the rhEPO we used a BLI biosensor. rhEPO was labeled with biotin and it was immobilised on a super streptavidin biosensor tip (SSA). The running buffer was phosphates 20 mM, pH 7.0, Tween 20, 0.05%. The reference biosensor tip was a SSA with immobilised biocitin. Peptides with the highest affinities were immobilised on agarose. All the peptide-agarose matrices showed affinity for rhEPO. Also, we evaluated the affinity of the peptide-agarose matrices for bovine seroalbumin, usually present in the culture supernatant. Those peptides with the highest selectivity were selected for future development of a rhEPO purification method.