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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

Author:
López, J., Lencina, F., Petruccelli, S., Marconi, P., Alvarez, M. A.
Source:
Plant cell, tissue, and organ culture 2010 v.103 no.3 pp. 307-314
ISSN:
0167-6857
Subject:
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
Abstract:
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.
Agid:
794751