Callahan Shellie M, Wonganan Piyanuch, Obenauer-Kutner Linda J, Sutjipto Suganto, Dekker Joseph D, Croyle Maria A
The University of Texas at Austin, College of Pharmacy, Division of Pharmaceutics, Austin, TX 78712-1074, USA.
J Virol Methods. 2008 Mar;148(1-2):132-45. doi: 10.1016/j.jviromet.2007.10.027. Epub 2007 Dec 21.
Inactivated viruses are important tools for vaccine development and gene transfer. 8-Methoxypsoralen (8-MOP) and long-wavelength ultraviolet irradiation (LWUVI) inactivates many viruses. Toxicity limits its use in animals and humans. Toxicological and photosensitizing properties of riboflavin make it suitable for virus inactivation in preparations for biological use. Viruses expressing beta-galactosidase were mixed with either 8-MOP (1.5mM) or riboflavin (50 microM) and exposed to LWUVI (365 nm) for 2 h. Virus activity was determined by limiting dilution. The half-life of the adenovirus preparation treated with 8-MOP was 8.28 ns(-1) and 36.5 ns(-1) after treatment with riboflavin. Despite the difference in half-life, both preparations were completely inactivated within 45 min. In contrast, the half-lives for adeno-associated virus (AAV) preparations were similar (63 ns(-1) 8-MOP vs. 67 ns(-1) riboflavin). Each AAV preparation was fully inactivated within 90 min. The half-life of lentivirus was 193.4 ns(-1) after treatment with 8-MOP and 208 ns(-1) after exposure to riboflavin. Virus treated with riboflavin was inactivated within 20 min. Virus exposed to 8-MOP was inactivated in 90 min. DNA and RNA viruses can be inactivated by riboflavin and LWUVI and used in physiological systems sensitive to other photochemicals.
灭活病毒是疫苗开发和基因转移的重要工具。8-甲氧基补骨脂素（8-MOP）和长波长紫外线照射（LWUVI）可灭活多种病毒。其毒性限制了它在动物和人类中的使用。核黄素的毒理学和光敏特性使其适用于生物制剂中的病毒灭活。将表达β-半乳糖苷酶的病毒与8-MOP（1.5mM）或核黄素（50μM）混合，并暴露于LWUVI（365nm）下2小时。通过有限稀释法测定病毒活性。用8-MOP处理的腺病毒制剂的半衰期为8.28ns(-1)，用核黄素处理后为36.5ns(-1)。尽管半衰期不同，但两种制剂在45分钟内均被完全灭活。相比之下，腺相关病毒（AAV）制剂的半衰期相似（8-MOP处理后为63ns(-1)，核黄素处理后为67ns(-1)）。每种AAV制剂在90分钟内均被完全灭活。慢病毒经8-MOP处理后的半衰期为193.4ns(-1)，暴露于核黄素后为208ns(-1)。用核黄素处理的病毒在20分钟内被灭活。暴露于8-MOP的病毒在90分钟内被灭活。DNA和RNA病毒可被核黄素和LWUVI灭活，并可用于对其他光化学物质敏感的生理系统。
