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

López, J., Lencina, F., Petruccelli, S., Marconi, P., Alvarez, M. A.
Plant cell, tissue, and organ culture 2010 v.103 no.3 pp. 307-314
Nicotiana tabacum, tobacco, cell suspension culture, biopharmaceuticals, molecular farming, recombinant antibodies, transgenic plants, endoplasmic reticulum, secretion, crop yield, osmotic pressure, mannitol, yields, culture media, dimethyl sulfoxide, adverse effects
We have established two transgenic cell suspension culture lines of Nicotiana tabacum that express the catalytic antibody 14D9 as a secretory product (sec-Ab) or as a KDEL-tagged product in the endoplasmic reticulum (Ab-KDEL), respectively. After 3 years of culture, the performance improved to a production level of 0.15 ± 0.03 μg ml⁻¹ on the seventh day of culture for the sec-Ab line and 0.48 ± 0.05 μg ml⁻¹ on the third day for Ab-KDEL line. Analysis of the effect of osmotic stress using mannitol (90 g l⁻¹) as an osmolite revealed that there was a 12-fold increase in antibody yield (1.96 ± 0.20 μg ml⁻¹) on the seventh day of culture in line sec-Ab and a fivefold increase (2.31 ± 0.18 μg ml⁻¹) on the seventh day for line Ab-KDEL. The concentration of the antibody in the culture medium was not significant. Dimethyl sulfoxide used as a permeabilizing agent was not effective in increasing 14D9 yield, but it did cause distinctive cell damage at all concentrations tested.