Influence of the KDEL signal, DMSO and mannitol on the production of the recombinant antibody 14D9 by long-term Nicotiana tabacum cell suspension culture
We have established two transgenic cell suspension culture lines of <i>Nicotiana tabacum</i> 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 performanc...
Guardado en:
| Autores principales: | , , , , |
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| Formato: | Articulo |
| Lenguaje: | Inglés |
| Publicado: |
2010
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| Materias: | |
| Acceso en línea: | http://sedici.unlp.edu.ar/handle/10915/133823 |
| Aporte de: |
| Sumario: | We have established two transgenic cell suspension culture lines of <i>Nicotiana tabacum</i> 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<sup>−1</sup> on the seventh day of culture for the sec-Ab line and 0.48 ± 0.05 μg ml<sup>−1</sup> on the third day for Ab-KDEL line. Analysis of the effect of osmotic stress using mannitol (90 g l<sup>−1</sup>) as an osmolite revealed that there was a 12-fold increase in antibody yield (1.96 ± 0.20 μg ml<sup>−1</sup>) on the seventh day of culture in line sec-Ab and a fivefold increase (2.31 ± 0.18 μg ml<sup>−1</sup>) 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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