Production of the recombinant antibody 14D9 by Nicotiana tabacum cell suspension cultures: influence of the KDEL signal, DMSO and mannitol. Establishment of improved lines by long –term in vitro culture

LÓPEZ, J; LENCINA, F; PETRUCCELLI, S; MARCONI, P AND ALVAREZ MA

PLANT CELL TISSUE AND ORGAN CULTURE

SPRINGER

Año: 2010

0167-6857

We have established transgenic cell suspension cultures of Nicotiana tabacum that express the catalytic antibody 14D9 as a secretory (Sec-Ab) or KDEL tagged version (Ab-KDEL). After three years in culture their performance improved until reaching a production level of 0.15 ± 0.03 ìg ml-1 on the 7th day of culture for sec-Ab line and of 0.48 ± 0.05 ìg ml-1 on the 3rd day for Ab-KDEL line. When we analyzed the effect of the osmotic stress by using mannitol 90 g l-1 as osmolite, the antibody yield showed a 12-fold increase (1.96 ± 0.20 ìg mL-1) on the 7th day of culture in line sec-Ab and a 5-fold increase (2.31 ± 0.18 ìg ml-1) on the 7th day for line Ab-KDEL. The antibody concentration into the culture medium was not significant. Dimethyl sulfoxide used as a permeabilising agent was not effective for increasing 14D9 yield; on the contrary it caused remarkable cell damage at all the concentrations tested.