The expression of the 14D9 catalytic antibody in suspended cells ofNicotiana tabacumcultures increased by the addition of protein stabilizers and by transference from erlenmeyer flasks to a 2-L bioreactor

What is it about?

The effect of two protein stabilizers (polyvinylpyrrolidone [PVP] and gelatine) on growth and 14D9 yield of Nicotiana tabacum cell suspension cultures (Ab-KDEL and sec-Ab) was analyzed. The addition of PVP at a concentration of 1.0 g/ L produced the highest total 14D9 yield (biomass + culture medium) in the Ab-KDEL line (4.82% total soluble protein [TSP]). With the addition of gelatine, the highest total 14D9 yield (2.48% TSP) was attained in the Ab-KDEL line at 5.0 g / L gelatine. When the Ab-KDEL suspended cells were cultured in a 2-L bioreactor, the highest 14D9 yield was 8.1% TSP at a 5% w/v inoculum size, which was the best 14D9 yield so far obtained in the platforms tested (E. coli, N. tabacum leaves and seeds, N. tabacum hairy roots, and cell suspension cultures).

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Why is it important?

The yield of a recombinant protein expressed in plant cell suspension of Nicotiana tabacum was increased by the addition of agents that protect the protein from degradation and also by scaling-up the culture to a bioreactor with mechanical agitation and stirring which increase the transfer of materia in the culture.