title: Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography.
abstract: Visceral leishmaniasis, a disease caused by Leishmania infantum chagasi, represents a major public health problem in many areas of the world. However, there is currently no vaccine for human use. The aim of this work was to purify the 503 antigen of Leishmania i. chagasi directly from unclarified Escherichia coli feedstock through expanded bed adsorption (EBA) chromatography. Batch experiments were performed to optimize the adsorption and elution conditions of the antigen onto a STREAMLINE Chelating resin using two central composite rotatable designs (CCRD). The results showed that the optimal binding conditions of the 503 antigen were pH 8.0 in the presence of 2.4 M NaCl. For the elution of the target protein, the optimized conditions included the presence of 600.0 mM imidazole. The adsorption isothermal data of the 503 antigen were fitted to the Langmuir adsorption isotherm. The EBA experiment successfully recovered 59.2% of the 503 antigen from the unclarified E. coli homogenate with a purification factor of 6.0.
authors.affiliation: Departamento de Engenharia Química, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil. Electronic address: fcsousa@eq.ufrn.br.
authors.forename: Francisco Caninde
authors.initials: FC
authors.lastname: de Sousa Junior
authors.affiliation: Departamento de Engenharia Química, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil; Centro de Desenvolvimento Sustentável do Semiárido, Universidade Federal de Campina Grande, Sumé, PB, Brazil.
authors.forename: Michelle Rossana Ferreira
authors.initials: MR
authors.lastname: Vaz
authors.affiliation: Departamento de Engenharia Química, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Carlos Eduardo
authors.initials: CE
authors.lastname: de Araújo Padilha
authors.affiliation: Departamento de Engenharia Química, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Abimaelle Silva
authors.initials: AS
authors.lastname: Chibério
authors.affiliation: Departamento de Biologia Celular e Genética, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Daniella Regina Arantes
authors.initials: DR
authors.lastname: Martins
authors.affiliation: Departamento de Engenharia Química, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Gorete Ribeiro
authors.initials: GR
authors.lastname: de Macedo
authors.affiliation: Departamento de Engenharia Química, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Everaldo Silvino
authors.initials: ES
authors.lastname: dos Santos
mesh_terms: D000327:Adsorption; D000953:Antigens, Protozoan / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D018314:Leishmania infantum; D016014:Linear Models; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D000953:Antigens, Protozoan; D011994:Recombinant Proteins
keywords: Expanded bed adsorption; Leishmania infantum chagasi; Recombinant protein purification; Unclarified bacterial homogenate; Visceral leishmaniasis