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Doctoral Thesis
DOI
https://doi.org/10.11606/T.87.2016.tde-26082016-091800
Document
Author
Full name
Dirce Sakauchi
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2016
Supervisor
Committee
Cianciarullo, Aurora Marques (President)
Almeida, Maria Elisabete Sbrogio de
Jorge, Soraia Attie Calil
Kubrusly, Flavia Saldanha
Ventura, Armando Morais
Title in Portuguese
Potencial vacinal de proteínas recombinantes do capsídeo de papilomavírus humano.
Keywords in Portuguese
Câncer
HPV16
L1L2
Papilomavírus humano
Vacina profilática
VLP
Abstract in Portuguese
O câncer cervical é a consequência mais séria da infecção por Papilomavírus humano - HPV, constituindo uma das principais causas de morte entre mulheres no mundo, remetendo a um importante desafio para a saúde pública mundial. Desenvolvemos a produção de VLPs contendo as proteínas L1 e L2 de HPV16, utilizando células epiteliais humanas 293-F em suspensão e em meio isento de soro fetal bovino. As células possuem metabolismo intenso, adequado para a expressão de proteínas heterólogas. Apresentam vantagens como a facilidade de cultivo, transfecção e ocorrência de processos como a glicosilação de proteínas, fosforilação, formação de pontes dissulfeto e outras modificações pós-traducionais essenciais para a função das proteínas, facilitando a produção similar às condições in vivo. O sistema de produção de VLPs L1L2 de HPV16 foi estabelecido de maneira eficiente, em células epiteliais humanas em suspensão e com resultados promissores, podendo contribuir para o desenvolvimento de uma vacina profilática de amplo espectro de proteção, com custo reduzido de produção.
Title in English
Vaccine potential of recombinant proteins of human papillomavirus capsid.
Keywords in English
Cancer
HPV16
Human papillomavirus
L1L2
Prophylactic vaccine
VLP
Abstract in English
Cervical cancer is the most serious consequence of infection by human papillomavirus - HPV, constituting one of the leading causes of death among women worldwide that points to a major challenge to global public health. We have developed the production of VLPs containing L1 and L2 proteins of HPV16, using human epithelial cells 293-F suspended in medium without fetal bovine serum. The cells have intense metabolism, suitable for expressing heterologous proteins. Present advantages such as ease of culture, transfection and occurrence of processes such as protein glycosylation, phosphorylation, formation of disulfide bonds and other essential post-translational modifications to protein function, facilitating the production similar to the conditions in vivo. The VLPs L1L2 of HPV16 production system was established efficiently in human epithelial cells in suspension and with promising results, which may contribute to the development of a prophylactic vaccine broad protection spectrum with reduced production cost.
 
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Publishing Date
2016-08-26
 
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