Preparation of human rabies vaccine in VERO cell culture using a microcarrier system.
1. The rabies virus (Pasteur PV strain) was propagated in VERO cells attached to microcarriers in a 3.7-1 bioreactor. Virus titers of about 10(6) LD50/ml were obtained regularly. 2. Ultrafiltration was efficient for concentrating the virus suspensions, and the sucrose gradient reduced the residual VERO cell DNA to acceptable levels (less than 50 pg/dose). The remaining cell DNA content was evaluated by dot-blot hybridization with a probe prepared with VERO cell DNA. 3. The final virus preparations were inactivated by B-propiolactone treatment, showed a potency higher than 2.5 IU/dose and protected mice experimentally infected intracerebrally with rabies virus (CVS-13.2). 4. This methodology for the production of a rabies vaccine for human use should be of interest to countries where high technology facilities are not available.
Mendonça
R Z
RZ
Instituto Butantan, São Paulo, Brasil.
Ioshimoto
L M
LM
Mendonça
R M
RM
De-Franco
M
M
Valentini
E J
EJ
Beçak
W
W
Raw
I
I
Pereira
C A
CA
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D004247:DNA / Q000032:analysis; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D009693:Nucleic Acid Hybridization; D011819:Rabies Vaccines / Q000276:immunology*; D011820:Rabies virus / Q000254:growth & development / Q000276:immunology*; D013997:Time Factors; D015164:Vaccines, Inactivated / Q000276:immunology; D014709:Vero Cells
D000914:Antibodies, Viral; D011819:Rabies Vaccines; D015164:Vaccines, Inactivated; D004247:DNA