High-Level Expression and Purification of Biologically Active Recombinant Pokeweed Antiviral Protein

摘要: Pokeweed antiviral protein (PAP) from the leaves of pokeweed plant, Phytolacca americana, is a naturally occurring single-chain ribosome-inactivating protein, which catalytically inactivates both prokaryotic and eukaryotic ribosomes. The therapeutic potential PAP has gained considerable interest in recent years due to clinical use native as active moiety immunoconjugates against cancer AIDS. limited inherent difficulties obtaining sufficient quantities homogenously pure preparation with minimal batch variability its natural source. Previous methods for expression recombinant yeast, transgenic plants Escherichia coli have resulted either unacceptably low yields or were too toxic host system. Here, we report successful strategy allows high level inclusion bodies E. coli. Purification refolded solubilized by size-exclusion chromatography yielded biologically (final yield: 10 12 mg/L). ribosome depurinating vitro N-glycosidase activity cellular anti-HIV comparable those PAP. This purification system makes it possible obtain homogenous carry out advanced trials. To our knowledge, this first large-scale