Binary ethylenimine as an inactivant for foot-and-mouth disease virus and its application for vaccine production

Abstract

Foot-and-mouth disease virus was inactivated with binary ethylenimine formed apart from or directly in the virus suspension by the cyclization of 2-bromoethylamine hydrobromide or 2-chloroethylamine hydrochloride under alkaline conditions. The inactivation rates with binary ethylenimine prepared apart from the virus suspension in dilute sodium hydroxide with either 2-bromoethylamine hydrobromide or 2-chlorethylamine hydrochloride were higher than with pure ethylenimine. When binary ethylenime was prepared directly in the virus suspension only 2-bromoethylamine hydrobromide gave acceptable inactivation rates. The reduced inactivation rates for binary ethylenimine directly prepared in the virus suspension are due to the different cyclization rates of 2-bromoethylamine hydrobromide and 2-chloroethylamine hydrochloride and to the interference of bicarbonate in the cyclization reaction. The complement fixing antigen of foot-and-mouth disease virus was not affected by binary ethylenimine inactivation. Vaccines prepared with foot-and-mouth disease virus inactivated by binary ethylenimine were comparable in their immunogenicity to vaccines prepared with ethylenimine or N-acetylethylenimine used as inactivants. Application of binary ethylenimine in the preparation of foot-and-mouth disease vaccines considerably reduces the potential danger associated with handling pure ethylenimine and other aziridines.

Heparan sulphate mediates swine vesicular disease virus attachment to the host cell

“…The virus was then concentrated by ultracentrifugation through 2 ml 20 % sucrose for 3 h at 110 000 g. For BIAcore analysis, the virus was purified on 10-35 % sucrose gradients as described by Seechurn et al (1990). SVDV was inactivated with 6 mM bromoethylimine (BEI) for 30 h at 28 uC (Bahnemann, 1975). Heparin, heparan sulphate (HS), chondroitin sulphate A, B and C, platelet factor 4 (PF4), and heparinases I and III were purchased from Sigma.…”

Neutralization of Foot-and-Mouth Disease Virus. I. Sensitization of the 140S Virion by Antibody Also Reactive with the 12S Protein Subunit

“…The A~2 BHK2 virus was inactivated with 3 mM-ethylenimine (BEI; Bahnemann, 1975) at 25 °C for 72 h, after which the reaction was stopped by addition of a molar excess of sodium thiosulphate (Brown & Cartwright, 1963). The BEI-inactivated virus was concentrated 100 times by precipitation with polyethylene glycol 6000 (PEG) and resuspended in barbital-glycine buffer containing 0.15 M-NaC1 .…”

Comparison of the immune response elicited by infectious and inactivated foot-and-mouth disease virus in mice

“…inoculation of 104 suckling mouse 50% lethal doses (SMLDso) of FMDV subtype O1 Campos (O1C) diluted in 0-5 ml of PBS per mouse. FMDV was inactivated with binary ethylenimine, according to Bahnemann (1975) and virus purification was performed as described by Bachrach et al (1964). Mice were immunized by i.p.…”