INVESTIGADORES
MARANI mariela Mirta
congresos y reuniones científicas
Título:
Bio-layer interferometry using optic-based biosensor for screening of affinity ligand for rhEPO
Autor/es:
MARTÍNEZ CERON MARÍA C.; MARANI MARIELA M.; ETCHEVERRYGARAY MARINA; TAULÉS MARTA; ALBERICIO FERNANDO; CASCONE OSVALDO; CAMPERI SILVIA A.
Lugar:
Puerto Madryn, Chubut, ARGENTINA
Reunión:
Congreso; SAIB XLVI Reunión anual (Sociedad Argentina de Investigación en Bioquímica y Biología Molecular); 2010
Resumen:
Bio-layer interferometry is used in optic-based biosensors (FortéBio 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 supernatants. Those peptides with the highest selectivity were selected for future development of a rhEPO purification method.