Abstract
The catalytic antibody 14D9 is an IgG1-like murine antibody that catalyzes the highly enantioselective protonation of enol-ethers. It was utilised as experimental model for studying the expression of an antibody in tobacco.
The ability of Nicotiana tabacum to express the whole antibody was confirmed. Also, in vitro cultures of N. tabacum were established and their ability to express the antibody was demonstrated. Attempts to improve the 14D9 initial yields were performed by the addition of a KDEL endoplasmic reticulum (ER) retention signal to the construct, therefore two N. tabacum lines were used in all the experiments, one with the secretory variant of the antibody (sec-Ab), and another one that retains the antibody in the ER (Ab-KDEL).
Other strategies tested were the optimization of plant growth regulator balance in the culture medium, and the addition of protein stabilizers, plant cell wall permeabilizers and osmotic agents. Besides, hairy root cultures were established and the conditions to express 14D9 were analyzed.
Comparing the yields obtained in all the platforms and system examined, we can conclude that hairy roots growing in Erlenmeyer flasks and Ab-KDEL cell suspension cultures growing in a 2-l bioreactor gave the highest 14D9 yields.
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Alvarez, M.A. (2014). The Antibody 14D9 as an Experimental Model for Molecular Farming . In: Plant Biotechnology for Health. Springer, Cham. https://doi.org/10.1007/978-3-319-05771-2_7
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DOI: https://doi.org/10.1007/978-3-319-05771-2_7
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