Transient expression of rabies virus G-glycoprotein using BHK-21 cells cultured in suspension

Autor(es)

Fernandez-Nunez, Eutimio Gustavo ^{[1, 2]} ; de Rezende, Alexandre Goncalves ^[3] ; Pradella Puglia, Ana Lia ^[3] ; Leme, Jaci ^[4] ; Boldorini, Vera Lucia Lopes ^[3] ; Caricati, Celso Pereira ^[4] ; Tonso, Aldo ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Lab Celulas Anim, Dept Engn Quim, Escola Politecn, BR-05508900 Sao Paulo, SP - Brazil

^[2] Univ Estadual Julio de Mesquita Filho, Dept Ciencias Biol, BR-19806900 Assis, SP - Brazil

^[3] Inst Butantan, Lab Imunol Viral, BR-05503900 Sao Paulo, SP - Brazil

^[4] Inst Butantan, Lab Especial Pesquisa & Desenvolvimento Imunol Ve, BR-05503900 Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Biotechnology Letters; v. 37, n. 6, p. 1153-1163, JUN 2015.
• Citações Web of Science: 0

Resumo

To assess the expression of rabies virus G-glycoprotein (RVGP) expression using Semliki Forest virus as a vector in combination with BHK-21 cells cultured in suspension. A multilevel factorial design was used to quantify effects of temperature (33-37 A degrees C), fresh medium addition after the viral adsorption step (100-200 % with respect to the initial cell suspension volume before infection) and harvest time (8-40 h) on RVGP production. Experimental runs were performed in 24-well cell culture plates at a multiplicity of infection (MOI) of 16. An additional experiment in spinner-flask was performed at MOI of 9, using the optimal conditions determined in cell culture plates. Values for temperature, fresh medium addition and harvest time of 33 A degrees C, 100 % and 16 h, respectively, ensured the optimal RVGP production in culture plates. The volumetric yield (239 ng ml(-1)) in these conditions was higher than that reported previously for adherent cell culture. In spinner-flasks, the volumetric yield was improved (559 ng ml(-1)). These results establish the basis for designing bioprocess to produce RVGP. (AU)

• Processo FAPESP: 10/52521-6 - Producao de glicoproteinas da raiva atraves do cultivo de celulas bhk-21 em biorreatores usando sistema de expressao semliki forest virus (sfv).
• Beneficiário: Eutimio Gustavo Fernández Núñez
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado