WOLMAN Federico Javier
congresos y reuniones científicas
Selective capture of recombinant bovine lactoferricin by affinity adsoption with triazinic dyes
NICOLÁS URTASUN; MARÍA FERNANDA BAIELI; OSVALDO CASCONE; FEDERICO J. WOLMAN; MARÍA V. MIRANDA
Congreso; SAIB 2012, 48 Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2012
Antimicrobial peptides (AMPs) are the first defense line against pathogens in many organisms. Bovine lactoferricin (Lfcin B) belongs to the AMPs family and has 25 amino acids with a net charge of +8 at physiological conditions. It is present in the amino terminal region of the bovine lactoferrin. Besides its antiviral, antifungal, antiparasitic, anticancer and antibacterial activity, Lfcin B has a synergistic effect with some conventional antibiotics. In this work, Lfcin B was expressed alone or as a fusion protein with a glutathione-S-transferase (Lfcin B-GST) using Baculovirus Expression System - Sf9 insect cells. When Lfcin B-GST was expressed and accumulated in cytoplasm or secreted to the culture supernatants, yields were 1.9 ± 0.4 and 1.1 ± 0.1 mg of Lfcin B/l, respectively. These values were 10 fold greater than Lfcin B expression without GST. For its recovery and purification, five triazinic dyes were screened using Surface Plasmon Resonance technology. The Yellow HE-4R and the Red HE-3B dyes showed the highest affinities for Lfcin B with Kd values of 1.1 ± 0.3 x 10-7 M and 1.6 ± 0.4 x 10-5 M, respectively. These dyes were immobilized on sepharose-4B matrix for further purification studies. The observed levels of Lfcin B-GST expression and its specific interaction with these low cost ligands are promising for the scaling up using lepidopteran larvae for industrial purposes.