Bio-Rad Sponsored Content

Yae Kurosawa,1 Maiko Saito,1 Daniel Yoshikawa,2 and Mark Snyder2 
1HOYA Technosurgical Corporation, Tokyo, Japan 
2Bio-Rad Laboratories, Inc., 2000 Alfred Nobel Drive, Hercules, CA, USA 

Mammalian Virus Purification Using Ceramic Hydroxyapatite 

Abstract

Conventional techniques for mammalian virus purification produce variable quality, quantity, and significant loss of particle infectivity. Here, we propose the chromatographic separation of viral particles of diverse sizes and from different families, such as dengue, Japanese encephalitis, influenza, mouse hepatitis, adenovirus, poliovirus, and feline calicivirus, using CHT™ Ceramic Hydroxyapatite Media (Resin). The separation of viral particles from impurities in at least one case was best observed on CHT Ceramic Hydroxyapatite Type II Media when compared to three other apatite media, suggesting the importance of determining which media works best for a specific virus. Additionally, factors such as adjusting the flow rate and gradient slope showed variable differences in the separation of viral particles on ceramic hydroxyapatite media.

Viruses can infect mammalian cells and cause diseases such as influenza, hepatitis, yellow fever, smallpox, and AIDS. Since some biotherapeutic products are produced using mammalian cell lines or plasma, the risk of viral contamination in these products is a concern and guidelines have been enforced to alleviate this risk. Chromatographic separation of viral particles from process intermediates is a key part of ensuring viral safety in biotherapeutics (ICH Expert Working Group 1999, Mbritz 2005). Additionally, purification of viral particles is used extensively in the study and characterization of these infectious agents. Understanding aspects of a virus, such as how it infects host cells, uses the host cells for reproduction, and evades the host immune system, aids scientists in determining how to use viruses for research and therapy. 

In order to study a virus, a pure, high-quality, infectious population is required. Conventional techniques for mammalian virus purification, for uses such as vaccine production or biological studies, can produce material of variable quality and quantity, often with significant loss of particle infectivity. 

In this paper, we report the use of ceramic hydroxyapatite media for purification of a wide variety of mammalian viruses. Chromatography using ceramic hydroxyapatite media is simple, easily scalable, and results in a concentrated preparation of highly active virus. 
 
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