title: Production of pharmaceutical-grade plasmids at high concentration and high supercoiled percentage.
abstract: The increased use of plasmid-based vaccines to replace their more challenging viral counterparts has increased the demand for high purity and high concentration plasmids. Here we report the production of plasmids encoding different transgenes for DNA vaccine candidates at gram scale with an integrated process consisting of batch fermentation and limited steps of purification. Plasmid products encoding for eight smallpox antigens that were combined into a bioterrorism DNA vaccine exhibited high purity with undetectable RNA, protein and endotoxin, concentration of up to 13.6mg/mL and supercoiled percentage of 94.5+/-1.1% after storage at -80 degrees C for over 1 year. The process has been scaled up for the cGMP manufacture of pharmaceutical-grade human papillomavirus and influenza DNA vaccines up to a 50g scale, also demonstrating high purity and high concentration.
authors.affiliation: VGX Pharmaceuticals, Inc., The Woodlands, TX 77381, United States.
authors.forename: Ying
authors.initials: Y
authors.lastname: Cai
authors.forename: Stephen
authors.initials: S
authors.lastname: Rodriguez
authors.forename: Ramu
authors.initials: R
authors.lastname: Rameswaran
authors.forename: Ruxandra
authors.initials: R
authors.lastname: Draghia-Akli
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Juba
authors.forename: Henry
authors.initials: H
authors.lastname: Hebel
mesh_terms: D004247:DNA / Q000096:biosynthesis* / Q000302:isolation & purification; D005285:Fermentation; D007252:Influenza Vaccines / Q000096:biosynthesis; D053918:Papillomavirus Vaccines / Q000096:biosynthesis; D010957:Plasmids; D011786:Quality Control; D012900:Smallpox Vaccine / Q000096:biosynthesis; D019444:Vaccines, DNA / Q000096:biosynthesis*
chemical_list: D007252:Influenza Vaccines; D053918:Papillomavirus Vaccines; D012900:Smallpox Vaccine; D019444:Vaccines, DNA; D004247:DNA