title: Purification of densonucleosis virus by tangential flow ultrafiltration.
abstract: Purification at commercial scale of viruses and virus vectors for gene therapy applications and viral vaccines is a major separations challenge. Tangential flow ultrafiltration has been developed for protein purification. Here tangential flow ultrafiltration of parvoviruses has been investigated. Because these virus particles are small (18-26 nm), removal of host cell proteins will be challenging. The results obtained here indicate that 30, 50, and 100 kDa membranes reject the virus particles, whereas 300 kDa membranes allow some virus particles to pass into the permeate. The decrease in permeate flux for the 300 kDa ultrafiltration membrane is much greater than for the 30, 50, and 100 kDa membranes, indicating possible entrapment of virus particle in the membrane pores. The permeate flux and level of protein rejection is strongly affected by the cell culture growth medium. The results indicate that when developing a new process, it is essential that the cell culture and purification operations be developed in parallel.
authors.affiliation: Institute of Biopharmaceutical Technology, University of Applied Sciences, Giessen-Friedberg, 35390 Giessen, Germany.
authors.forename: David L
authors.initials: DL
authors.lastname: Grzenia
authors.forename: Jonathan O
authors.initials: JO
authors.lastname: Carlson
authors.forename: Peter
authors.initials: P
authors.lastname: Czermak
authors.forename: Binbing
authors.initials: B
authors.lastname: Han
authors.forename: Rachel K
authors.initials: RK
authors.lastname: Specht
authors.forename: S Ranil
authors.initials: SR
authors.lastname: Wickramasinghe
mesh_terms: D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D003470:Culture Media / Q000378:metabolism; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D019223:Densovirinae / Q000737:chemistry* / Q000302:isolation & purification*; D015316:Genetic Therapy / Q000295:instrumentation; D008567:Membranes, Artificial; D016133:Polymerase Chain Reaction; D011312:Pressure; D014462:Ultrafiltration; D014867:Water / Q000737:chemistry
chemical_list: D003470:Culture Media; D016895:Culture Media, Serum-Free; D008567:Membranes, Artificial; D014867:Water