| Autor: |
Sánchez ‐ Arreola, Pamela B., López ‐ Uriarte, Salvador, Marichal ‐ Gallardo, Pável A., González ‐ Vázquez, Juan C., Pérez ‐ Chavarría, Roberto, Soto ‐ Vázquez, Pedro, López ‐ Pacheco, Felipe, Ramírez ‐ Medrano, Alicia, Rocha ‐ Pizaña, María R., Álvarez, Mario M. |
| Předmět: |
|
| Zdroj: |
Biotechnology Progress; Jul2013, Vol. 29 Issue 4, p896-908, 13p, 2 Diagrams, 3 Charts, 6 Graphs |
| Abstrakt: |
The current commercial system for influenza vaccine production depends on the culture of virus in embryonated eggs-a strategy that is both costly and poorly scalable. Consequently, a sudden pandemic event with a demand for millions of vaccine doses in a short time could readily overwhelm the available world production capacity. In this communication, we present a process that uses Escherichia coli for scalable production of recombinant vaccine candidates against influenza. A monomeric and a dimeric fragment of hemagglutinin of the influenza A H1N1/2009 virus were successfully expressed in a BL21 (DE3) pLysS variety of C41 E. coli. We present results from batch processes where induction is made with isopropyl thiogalactoside and from fed-batch experiments where expression is induced using lactose/glucose pulses. Concentrations in the range of 1.188-0.605 g/L of recombinant protein were observed in 2-L stirred tank bioreactors. The genetic construct included an N-terminal histidine tag sequence that facilitated recovery, purification, and proper refolding of the vaccine candidate by affinity chromatography in columns loaded with Ni+2. The proteins produced by this strategy selectively and specifically recognizes antibodies from patients diagnosed as positive to influenza A H1N1/2009. Overall protein recovery yields between 30.0 and 34.7% were typically observed. Based on these yields, a production of 4.6 × 103 doses L−3 day−1 is feasible. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:896-908, 2013 [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
|
Plný text