Recovery and purification of recombinant 503 antigen of Leishmania infantum chagasi using expanded bed adsorption chromatography

• Autor(a) principal: Santos, Everaldo Silvino dos
• Data de Publicação: 2015
• Outros Autores: Sousa Junior, Francisco Caninde de, Vaz, Michelle Rossana Ferreira, Padilha, Carlos Eduardo de Araújo, Chibério, Abimaelle Silva, Martins, Daniella Regina Arantes, Macedo, Gorete Ribeiro de
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UFRN
• Texto Completo: https://repositorio.ufrn.br/handle/123456789/32482
• topic: Expanded bed adsorption; Leishmania infantum chagasi; Recombinant protein purification; Unclarified bacterial homogenate; Visceral leishmaniasis

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 STREAMLINETM Chelating resin using two central composite rotatable designs (CCRD). The results showed that the optimal binding con- ditions 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