REDBIO México 2010

Folio: 120
Tipo de presentación: Cartel
Tema: Otro

Influence of the KDEL signal and mannitol on the production of the recombinant antibody 14D9 by long-term Nicotiana tabacum cell suspension culture

Julieta López (1), Patricia Marconi (1), María Alejandra Alvarez * (1).
* Autor para correspondencia (Corresponding author) malvarez@conicet.gov.ar

1- Dpto Biotecnología Vegetal-Instituto de Ciencia y Tecnología Dr. César Milstein - CONICET. Saladillo 2468 14440 Ciudad de Buenos Aires, Argentina

RESUMEN (ABSTRACT)
"The ability of plant systems to produce complex recombinant proteins, such as antibodies, has been already demonstrated. The advantages of in vitro plant systems to produce biopharmaceuticals are, among others, their capacity to manage post-translational modifications, their culture medium simplicity, the absence of animal or human pathogenes and the feasibility of performing the productive process under GMP and GLP. We have already demonstrated the ability of Nicotiana tabacum whole plants and hairy roots to express the IgG1-like murine antibody 14D9. In this work we are reporting the establishment of N. tabacum cell suspensión cultures to produce 14D9. Also, we have analyzed the influence of the ER retention signal KDEL on the antibody yield. The antibody content increased approximately 5-fold in the line with the KDEL signal (Ab-KDEL) respect to the line without KDEL (sec-Ab). One remarkable finding of this work is that we have improved the culture performance by periodical subcultures to optimized medium during 3 years. Higher antibody levels (3-fold increase) and a shortened lag phase were attained which is preferable from a biotechnological point of view. On the other hand, by inducing an osmotic stress with mannitol (90 g l-1) we have obtained a 12-fold antibody increase."