Abstract
Electron beam (eBeam) inactivation of pathogens is a commercially proven technology in multiple industries. While commonly used in a variety of decontamination processes, this technology can be considered relatively new to the pharmaceutical industry. Rotavirus is the leading cause of severe gastroenteritis among infants, children, and at-risk adults. Infections are more severe in developing countries where access to health care, clean food, and water is limited. Passive immunization using orally administered egg yolk antibodies (chicken IgY) is proven for prophylaxis and therapy of viral diarrhea, owing to the stability of avian IgY in the harsh gut environment. Since preservation of viral antigenicity is critical for successful antibody production, the aim of this study was to demonstrate the effective use of electron beam irradiation as a method of pathogen inactivation to produce rotavirus-specific neutralizing egg yolk antibodies. White leghorn hens were immunized with the eBeam-inactivated viruses every 2 weeks until serum antibody titers peaked. The relative antigenicity of eBeam-inactivated Wa G1P[8] human rotavirus (HRV) was compared to live virus, thermally, and chemically inactivated virus preparations. Using a sandwich ELISA (with antibodies against recombinant VP8 for capture and detection of HRV), the live virus was as expected, most immunoreactive. The eBeam-inactivated HRV’s antigenicity was better preserved when compared to thermally and chemically inactivated viruses. Additionally, both egg yolk antibodies and serum-derived IgY were effective at neutralizing HRV in vitro. Electron beam inactivation is a suitable method for the inactivation of HRV and other enteric viruses for use in both passive and active immunization strategies.
Highlights
Routine pathogen inactivation methods in the pharmaceutical industry include chemical and thermal treatments
This study outlines the use of eBeam inactivation to produce high-titer neutralizing human rotavirus (HRV)-specific egg yolk antibodies and analyzes their potential to prevent infection in vitro
Electrons generated from High energy eBeam (HEEB) equipment were utilized to create extensive breaks in the RNA genome
Summary
Routine pathogen inactivation methods in the pharmaceutical industry include chemical and thermal treatments. Microbial inactivation results from both direct damage to the nucleic acids by electrons or indirectly from high reactive radiolytic species produced by the radiolysis of water molecules by the energetic electrons. Both single and double strand breaks can be generated rendering the organism inactive. Studies in our laboratories have shown that bacterial cells when inactivated retain their metabolism for specific periods of time in a state termed, Metabolically Active yet Non-culturable (MAyNC) [4] This technology is used commercially for different applications [2, 3], but its use for pathogen inactivation in pharmaceutical development is relatively new. Previous studies have compared electron beam irradiation to traditional methods of pathogen inactivation and concluded that eBeam is an effective alternative [2, 3, 5]
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