Production and purification of a recombinant fragment of pneumococcal surface protein A clade 3 (PspA3) from Streptococcus pneumoniae in Escherichia coli. / Produção e purificação de um fragmento recombinante da proteína A de superfície do clado 3 (PspA3) de Streptococcus pneumoniae  em Escherichia coli.

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

The pneumococcal surface protein A (PspA) is indispensable for virulence of S. pneumoniae and it was the first choice as carrier for a new conjugated vaccine against S.pneumoniae. Hence, the purpose of this work was to develop an industrial production and purification process of a recombinant fragment PspA clade 3 (rfPspA3) in E. coli. Fed-batch cultivations in 5 L bioreactors with defined medium were carried out using glucose or glycerol as carbon sources. It was obtained 62 g/L of dry cell weight and 3 g/L of rfPspA3. Cells were disrupted with 96.7% of efficiency by high pressure continuous homogenizer. Centrifugation was defined for the clarification step. The sequence with Q- followed by IMAC-Sepharose yielded the best purity and recovery of rfPspA3 (81 and 70%, respectively). Cation exchange was chosen for the last chromatography. In conclusion, an industrial production and purification process was developed and rfPspA3 was obtained with high purity (90%).

ASSUNTO(S)

alta densidade celular liquid chromatography streptococcus pneumoniae high cell density escherichia coli cromatografia líquida purificação de proteína recombinante clarificação escherichia coli recombinant protein purification clarification streptococcus pneumoniae

ACESSO AO ARTIGO

Documentos Relacionados