Influence of the KDEL signal,DMSO and mannitol on the production of the recombinant antibody 14D9 by long-term <Emphasis Type="Italic">Nicotiana tabacum</Emphasis> cell suspension culture |
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Authors: | J López F Lencina S Petruccelli P Marconi M A Alvarez |
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Institution: | 1.Instituto de Ciencia y Tecnología Dr. César Milstein, CONICET-Fundación Pablo Cassará,Ciudad de Buenos Aires,Argentina;2.Facultad de Ciencias Exactas, Universidad Nacional de la Plata,La Plata,Argentina |
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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. |
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