Controlled Inactivation of Recombinant Viruses with Vitamin B2

摘要

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 (1.5 mM) or riboflavin (50 μM) and exposed to LWUVI (365 nm) for 2 hours. Virus activity was determined by limiting dilution. The half-life of the adenovirus preparation treated with 8-MOP was 8.28 nanoseconds⁻¹ (ns⁻¹) and 36.5 ns⁻¹ after treatment with riboflavin. Despite the difference in half-life, both preparations were completely inactivated within 45 minutes. 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 minutes. 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 minutes. Virus exposed to 8-MOP was inactivated in 90 minutes. DNA and RNA viruses can be inactivated by riboflavin and LWUVI and used in physiological systems sensitive to other photochemicals.