The Effect of Benzonase Endonuclease Addition to Purification of Sabin Poliovirus Type 3

19-9-SS-Merck-Cover-229x300.pngDuring production of vaccines and viral vectors, the size and quantity of extracellular nucleic acids must be reduced using endonuclease enzymes. Merck/MilliporeSigma’s proprietary Benzonase endonuclease is a genetically engineered nuclease derived from the Gram-negative bacteria Serratia marcescens. It attacks and degrades all forms of DNA and RNA. It is manufactured under good manufacturing practice (GMP) conditions and has a drug master file (MDF) in place with the US Food and Drug Administration (FDA), which can be cited in regulatory filings. In this supplier-side article, authors from Merck/MilliporeSigma summarize the benefits of incorporating Benzonase endonuclease in a Sabin poliovirus type 3 production process.

Specifically, the authors examine the amount of Benzonase endonuclease required to degrade Vero cell DNA and describe its effect on clarification-filter capacity. Their study also explores the endonuclease’s effects on Vero-cell DNA levels following tangential-flow filtration (TFF), then suggests improvements to concentration using TFF. The authors ultimately show how the endonuclease product can reduce Vero DNA levels to <10 ng/dose and to fragments <200 base pairs (bp) in length, in compliance with World Health Organization (WHO) and FDA guidelines.

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The life science business of Merck, KGaA, Darmstadt, Germany, operates as MilliporeSigma in the United States and Canada.

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