Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo

• Ano de defesa: 2020
• Autor(a) principal: Soares, Gabriela de Fátima da Silva [UNIFESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo
• Palavras-chave em Português: Hibridoma; Cultura de células; Anticorpo monoclonal; Soro fetal bovino; Nutridoma
• Link de acesso: https://hdl.handle.net/11600/63864

Monoclonal antibodies (mAbs) are molecules involved in multiple applications, ranging from use as reagents to diagnostic and therapeutic approaches. Once obtained, mAbs can be produced in vivo, by inducing ascites, as well as in vitro, by culturing the secretory hybridoma cells. In view of changes in the laws that regulate the use of animals in research, the in vivo production method has become questionable. However, the production of mAbs in vitro is not trivial. In order to replace or reduce the use of animals for this purpose, we present the present work that sought to evaluate possibilities of mAb production in vitro. For this, we use hybridomas selected based on the degree of ease in the cultivation and production of mAb, under the conditions usually practiced. Hybridomas 1F5H2 (good secretory), 5.G8 (bad secretory) and 10. D7 (best secretory) were used. The three hybridomas were studied in order to allow the comparison between in vivo and in vitro production methods regarding cell proliferation under different conditions of culture medium supplementation - 10% conventional SFB, 10% low IgG SBF and 1% Nutridoma- , as to the secretory antibody capacity of hybridomas, as well as the cost, time and volume required to produce reasonable amounts of antibodies. The cells showed no significant difference regarding the proliferative profile in the different conditions of supplementation of the culture medium. However, the 5.G8 hybridoma exhibited a higher proliferative rate when compared to the other two hybridomas, showing that the hybridoma that presented the worst performance in in vivo production, performed better in vitro and that each strain has its own characteristics. In addition, we saw that no in vitro production method, evaluated for the three hybridoma strains studied, exceeded the profitability of the in vivo method. In vitro production has always involved longer times to obtain the same amount of antibody. Supplementation of medium with Nutridoma for hybridoma culture proved to be an alternative for culture in serum-free medium. The implementation of an in vitro method alternative to the use of animals will require investments and will depend on a demand that sustains a favorable cost / benefit ratio.