Monday, November 9, 2009
P30

Production of a recombinant fragment of pneumococcal surface protein A from clade 1 (rfPspA1) in fed-batch cultivation of Escherichia coli

Viviane M. Gonçalves1, Antonio J. G. Cruz2, Roberto de Campos Giordano2, Martha M. Tanizaki1, José G. C. Pradella3, and Joaquin Cabrera-Crespo1. (1) Centro de Biotecnologia, Instituto Butantan, Av Vital Brasil, 1500, São Paulo, Brazil, (2) Chemical Engineering Department, Universidade Federal de São Carlos-UFSCar, Washington Luiz, Km 235, São Carlos, Brazil, (3) Centro de Tecnologia do Bioetanol, Caixa Postal 6170, Campinas, Brazil

Streptococcus pneumoniae is an important pathogenic bacterium that causes pneumonia, meningitis, otitis media and bacteremia and presents more than 90 serotypes. New conjugated vaccines against this microorganism are being developed using pneumococcal proteins as carriers. The pneumococcal surface protein A (PspA) was the first choice as carrier because it is indispensable for virulence of S. pneumoniae and may induce higher protection and cross reaction between serotypes. Cultivation to produce recombinant protein at high cell density (HCD) is essential for vaccine development, because the higher amount of protein is produced; the lower is the cost of vaccine. The present work aimed to develop a production process of a recombinant fragment of pneumococcal surface protein A from clade 1 (rfPspA1) and reach high cell density in chemically defined medium. Four production protocols were evaluated using pre-defined specific flow rates for exponential feeding of substrate limiting growth in fed-batch phase and the impact of reducing the concentration of IPTG, expensive and toxic reagent, by using lactose on rfPspA1 production was investigated. The higher amount of rfPspA1, ~2.0g/L, was obtained when glucose was fed at 0.05h-1 during the induction phase with 0.5mM IPTG and 20g/L lactose.

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