OBTENTION OF RABIES ANTIGEN THROUGH BHK21 CELLS ADHERED TO MICROCARRIERS

Four rabies antigen batches were produced from virus suspensions resulting from BHK21 cells adhered to microcarriers (Cytodex 1), inoculated and cultured in a bioreactor. In parallel the methodology of production of rabies virus through cultures of BHK21 cells in monolayers in bottles was used. The...

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Bibliographic Details
Published in:Revista do Instituto de Medicina Tropical de São Paulo
Main Authors: Neuza M. FRAZATTI GALLINA, Rosana de LIMA PAOLI, Izilda A. FRANCISCO, Gislaine de CASTRO GARCIA, Regina M. MOURÃO FUCHES
Format: Article in Journal/Newspaper
Language:English
Published: Universidade de São Paulo (USP) 1998
Subjects:
Online Access:https://doi.org/10.1590/S0036-46651998000500005
https://doaj.org/article/b7fa4630dffe4952a6db1d1513ac65d6
Description
Summary:Four rabies antigen batches were produced from virus suspensions resulting from BHK21 cells adhered to microcarriers (Cytodex 1), inoculated and cultured in a bioreactor. In parallel the methodology of production of rabies virus through cultures of BHK21 cells in monolayers in bottles was used. The results obtained showed that infecting titles were 106.69 DL50/mL and 107.28 DL50/mL for suspensions cultured in bottles and in the bioreactor, respectively. The viral suspension volumes collected were on average 11,900 per batch from the bioreactor and 800mL per bottle. Ten horses were immunized with the antigen produced in the bioreactor. The means of antirabies antibody titers found were 240 and 212 IU/mL after the initial and the first booster doses, respectively. Rabies antigen with satisfactory infecting titers can be obtained on a large scale by culturing in a bioreactor inoculated BHK21 cells adhered to microcarriers. Foram produzidos quatro lotes de antígeno rábico a partir de suspensões de vírus resultantes de células BHK21 infectadas, aderidas a microcarregadores do tipo Cytodex 1 e cultivadas em biorreator. Em paralelo foi utilizada a metodologia de produção de vírus rábico com células BHK21 em monocamadas, contidas em garrafas de 350cm2. Os resultados encontrados demonstraram que os títulos infectantes foram de 106.69 DL50/mL para as suspensões virais obtidas em garrafas e 107.28 DL50/mL para as do biorreator. Os volumes das suspensões virais colhidas foram, em média de 11.900mL por lote do biorreator e 800mL por garrafa . Com o antígeno produzido no biorreator foram imunizados 10 cavalos. As médias dos títulos de anticorpos anti-rábicos encontrados no soros destes animais foram de 240 e 212 UI/mL, respectivamente após a base e o primeiro reforço. Através da infecção de células BHK21 aderidas a microcarregadores e cultivadas em biorreator, pode-se obter antígeno rábico em larga escala e com títulos infectantes satisfatórios.