Monday, February 13, 2012

UV-C versus small, non-enveloped viruses

Small, non-enveloped viruses (especially the circoviruses, parvoviruses, picornaviruses, caliciviruses, and polyomaviruses) and bacteriophage with similar characteristics represent a special challenge to the biologics industry.
In fact, contamination events have occurred with each of these virus families; in some cases more than once. Within the Circoviridae, the primary concern has been the porcine circovirus discovered in rotavirus vaccine. At least four contamination events involving the parvovirus, mouse minute virus, have been reported. The calicivirus vesivirus 2117 has contaminated biologics manufacturing processes on at least two occasions. The picornaviruses are a threat which has yet to be realized in the manufacturing context; but a viral safety test conducted for a biologics cell bank was the scene of a contamination with equine rhinitis A virus. The polyomavirus SV40 was found to have survived the formaldehyde inactivation used in the preparation of polio vaccines and therefore made its way into the doses of vaccine delivered to millions of individuals during the decade between 1955 and 1965 (being born in 1952, it is possible that the author received such tainted vaccine!).
The contamination events described above are, at least in part, a reflection of the ability of these viruses to withstand conditions that would lead to inactivation of other types of viruses. Inactivation strategies that are typically employed for viral safety include chemical (low pH, high pH, disinfectants, solvents, detergents, etc.) and physical (heat, irradiation, pressure, etc.) means. In a previous posting, the efficacy of gamma irradiation for inactivating various types of viruses within frozen animal serum was discussed. The use of UV-A in combination with riboflavin has also been discussed previously.
It is perhaps fortunate that the efficacies of different inactivation approaches are in many cases complementary. For instance, it appears that the efficacy of gamma irradiation for inactivation of viruses decreases as viral particle size decreases (although this relationship is not strictly linear). The outcome of this is that gamma irradiation does not appear to be particularly effective for inactivating small, non enveloped viruses from the circovirus, parvovirus, and polyomavirus families. On the other hand, ultraviolet radiation in the C range (254 nm is the most commonly employed wavelength) appears to be more effective for the inactivation of smaller viruses than for larger viruses (or bacteria).
The table below is a compilation of the UV-C inactivation constants (K, defined as the log10 reduction in titer per mJ/cm2 fluency) for various families of small, non-enveloped viruses. These K values represent inactivation of the viruses in a variety of matrices, ranging from water to protein-containing matrices such as albumin or complete culture medium). These results suggest that UV-C irradiation may be a viable approach for inactivating many of these problem viruses in raw materials or process intermediates used in biologics manufacture.

references for K values: * Lytle and Sagripanti 2005, ¶ Kowalski et al. 2009; † Maier 2007. The other values are the mean K values assembled by the author from the inactivation literature.
The one exception appears to be the polyomaviruses, which appear to be relatively resistant to the inactivating effects both of gamma irradiation and UV-C irradiation. This family of viruses may best be inactivated using high-temperature short-time treatment (HTST), though the efficacy of this approach for the polyomaviruses has yet to be demonstrated empirically.