Studies on the efficiency of measles virus antigen production using VERO cell culture in a microcarrier system.
1. A large amount of antigen is required to conduct seroepidemiologic surveys of measles. Thus, a process to obtain measles virus antigen using a bioreactor was standardized. 2. The virus was grown in a 3.7-1 culture of VERO cells using a Celligen cell culture system containing 2 mg/ml of microcarriers (cytodex I) at 37 degrees C and 60 rpm. The cultures infected with 0.5 m.o.i. of measles virus were harvested after the appearance of the cytopathic effect. The virus suspension was clarified and concentrated by ultracentrifugation. Intracellular and extracellular virus titers were determined by hemagglutination (HA) and by induction of a cytopathic effect in cell culture (TCID50). 3. Intracellular virus presented 5-7 x 10(6) TCID50/0.1 ml, HA activity per 50 microliters equal to 32, with a total HA activity of 4,480 HA units (HAU) and specific activity of 116 HAU/mg. In the concentrated supernatants, the HA titer of extracellular virus was 64, with a total HA activity of 1,024 HAU and a specific activity of 1,600 HAU/mg. 4. The antigen obtained was suitable for the detection of antibodies against measles virus in assays such as ELISA and DOT-ELISA (using 1 micrograms/well to ELISA and 2 micrograms/DOT). 5. The microcarrier system produced antigen sufficient for 26 ELISAs/ml compared to 5.7 ELISAs/ml obtained for the static culture system.
Mendonça
R Z
RZ
Laboratório de Immunologia Viral, Instituto Butantan, São Paulo, Brasil.
Vaz-de-Lima
L R
LR
Oliveira
M I
MI
Pereira
C A
CA
Hoshino-Shimizu
S
S
D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000276:immunology; D002522:Chlorocebus aethiops; D004797:Enzyme-Linked Immunosorbent Assay; D006386:Hemagglutination Tests; D006801:Humans; D008459:Measles virus / Q000254:growth & development* / Q000276:immunology; D014461:Ultracentrifugation; D014709:Vero Cells; D014776:Virus Cultivation
D000956:Antigens, Viral