Purification of Densonucleosis Virus by Tangential Flow Ultrafiltration

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• 作者：David L. Grzenia ; Jonathan O. Carlson ; Peter Czermak ; Binbing Han ; Rachel K. Specht ; S. Ranil Wickramasinghe
• 刊名：Biotechnology Progress
• 出版年：2006
• 出版时间：October 2006
• 年：2006
• 卷：22
• 期：5
• 页码：1346 - 1353
• 全文大小：335K
• 年卷期：v.22,no.5(October 2006)
• ISSN：1520-6033

文摘

Purification at commercial scale of viruses and virus vectors for gene therapy applications andviral vaccines is a major separations challenge. Tangential flow ultrafiltration has been developedfor 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 bechallenging. The results obtained here indicate that 30, 50, and 100 kDa membranes reject thevirus 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 thanfor the 30, 50, and 100 kDa membranes, indicating possible entrapment of virus particle in themembrane pores. The permeate flux and level of protein rejection is strongly affected by thecell culture growth medium. The results indicate that when developing a new process, it isessential that the cell culture and purification operations be developed in parallel.