Very fast (and safe) inactivation of foot-and-mouth disease virus and enteroviruses by a combination of binary ethyleneimine and formaldehyde.
For FMD vaccine production, inactivation of the FMD virus is the most critical step. Formerly, from 1940 onwards, the virus was inactivated with formaldehyde. This inactivation was relatively slow, about 0.2 - 0.3 log 10 per hour. Because formaldehyde not only reacts with the virus produced but with many other components in the medium, such as proteins and amino acids, its concentration can become rate-limiting and inactivation plots may show tailing-off, resulting in residual infectivity. Many of the bad stories of post-vaccination outbreaks date back to the use of formaldehyde-inactivated vaccines (e.g. the outbreaks in France in 1981 and in Eastern Germany causing the Danish outbreak in 1982). Much faster and safer inactivation was obtained with aziridines and in the 1980s binary ethyleneimine (BEI) was introduced in practically all vaccine production laboratories. If inactivation plots are made of every production batch, as is now required by the European Pharmacopoeia, and these plots show proper inactivation rates, vaccines can considered to be completely safe. Under optimal conditions, inactivation rates are in the range of 0.5 - 1.0 log 10 per hour. In general, the inactivation takes 40-48 hours,which will guarantee complete inactivation of all virus particles in a batch. Since formaldehyde (FA), the 'classical' inactivating agent, inactivates at a rate of 0.3 logs per hour only, a significant contribution of FA to the inactivation of BEI can hardly be expected. However, here it is shown that FA added during the BEI-inactivation process strongly augments inactivation rates with a hundred to thousand-times (to 2.5-3.5 logs per hour). This will enable inactivation during a working day or just overnight with even higher safety levels of the vaccines. Also, it is known that formaldehyde cross-links viral proteins which will stabilise the antigen. The short inactivation times will limit proteolytic destruction of 146 S antigen and increase antigen yields. It is expected that by the cross-linking activity of FA the stability of the antigen (and of vaccines) and the endurance of the immune response will be favourably influenced.
Barteling
S J
SJ
Simbar Consultancy, Amsterdam, The Netherlands. s.barteling@chello.nl
Cassim
N I
NI
D000818:Animals; D000956:Antigens, Viral / Q000276:immunology*; D001388:Aziridines / Q000494:pharmacology; D002417:Cattle; D002418:Cattle Diseases / Q000517:prevention & control*; D004306:Dose-Response Relationship, Immunologic; D005536:Foot-and-Mouth Disease / Q000517:prevention & control*; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D005557:Formaldehyde / Q000494:pharmacology; D009500:Neutralization Tests; D016896:Treatment Outcome; D014765:Viral Vaccines / Q000276:immunology*
D000956:Antigens, Viral; D001388:Aziridines; D014765:Viral Vaccines; D005557:Formaldehyde; C033132:aziridine