GIULIETTI Ana Maria
congresos y reuniones científicas
Production of a functional catalytic antibody 14D9 by transformed Nicotiana tabacum hairy root cultures
GIULIETTI, ANA MARÍA
Conferencia; 12th International Biotechnology Symposium; 2004
Plant tissue cultures are an alternative means for producing valuable proteins such as antibodies, enzymes, vaccines and therapeutic agents. We have expressed the catalytic antibody 14D9 (an IgG1murine-type) in hairy root cultures. We studied the expression level in the biomass and the culture medium during 30 days of culture. At 25 days of culture the final biomass were 0.521 g Fresh Weight / flask in wild type and transgenic hairy roots respectively. With respect to the biomass total antibody amount it was 4% of the Total Protein Soluble (TPS). Also the influence of the KDEL signal peptide for retention in the endoplasmic reticulum (RE) was analyzed. In this case the final biomass obtained was 2.290 g Fresh Weight / flask and the total antibody was 6% TPS. No antibody level nor protease activity were detected on the culture medium with the methodology used. In addition the influence of protein stabilizing agents (PVP, DMSO, gelatin) was studied. DMSO at a concentration of 2.8% v/v showed a positive influence with an increase of 2% in the 14D9 level with respect to non-treated cultures.