Scalable purification of Bacillus anthracis protective antigen from Escherichia coli |
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Authors: | Gwinn William Zhang Mei Mon Sandii Sampey Darryl Zukauskas David Kassebaum Corby Zmuda Jonathan F Tsai Amos Laird Michael W |
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Institution: | Human Genome Sciences, Inc., Rockville, MD 20850, USA. |
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Abstract: | The anthrax toxin consists of three proteins, protective antigen (PA), lethal factor, and edema factor that are produced by the Gram-positive bacterium, Bacillus anthracis. Current vaccines against anthrax use PA as their primary component. In this study, we developed a scalable process to produce and purify multi-gram quantities of highly pure, recombinant PA (rPA) from Escherichia coli. The rPA protein was produced in a 50-L fermentor and purified to >99% purity using anion-exchange, hydrophobic interaction, and hydroxyapatite chromatography. The final yield of purified rPA from medium cell density fermentations resulted in approximately 2.7 g of rPA per kg of cell paste (approximately 270 mg/L) of highly pure, biologically active rPA protein. The results presented here exhibit the ability to generate multi-gram quantities of rPA from E. coli that may be used for the development of new anthrax vaccines and anthrax therapeutics. |
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