TY - JOUR
T1 - Production of human secreted alkaline phosphatase in suspension and immobilization cultures of tobacco NT1 cell
AU - Nakashima, Kazunori
AU - Shibasaki-Kitakawa, Naomi
AU - Miyamoto, Takuya
AU - Kubo, Masaki
AU - Yonemoto, Toshikuni
AU - Shuler, Michael L.
PY - 2013/8/15
Y1 - 2013/8/15
N2 - The recombinant protein production by a plant cell culture offers several advantages including product safety and ease of the purification of secreted proteins and the capability of plants to correctly fold complex proteins. However, low productivity is generally a limitation of this system, caused by the denaturation and degradation of the secreted proteins due to proteolytic enzyme protease in the medium. In this study, we demonstrated that the transgenic tobacco NT1 cells were immobilized in Ca2+-alginate gel beads coated with a cell-free gel film, which significantly improved the productivity of the recombinant human secreted alkaline phosphatase (SEAP), about 33 times larger than that in the suspension culture. The cells immobilized in the beads were protected from hydrodynamic stress during cultivation and hence did not release the endogenous protease, which caused damage to the secreted SEAP. The immobilization culture without cell leakage could be quite effective for the production of recombinant protein.
AB - The recombinant protein production by a plant cell culture offers several advantages including product safety and ease of the purification of secreted proteins and the capability of plants to correctly fold complex proteins. However, low productivity is generally a limitation of this system, caused by the denaturation and degradation of the secreted proteins due to proteolytic enzyme protease in the medium. In this study, we demonstrated that the transgenic tobacco NT1 cells were immobilized in Ca2+-alginate gel beads coated with a cell-free gel film, which significantly improved the productivity of the recombinant human secreted alkaline phosphatase (SEAP), about 33 times larger than that in the suspension culture. The cells immobilized in the beads were protected from hydrodynamic stress during cultivation and hence did not release the endogenous protease, which caused damage to the secreted SEAP. The immobilization culture without cell leakage could be quite effective for the production of recombinant protein.
KW - Coated gel beads
KW - Immobilization culture
KW - Plant cells
KW - Protein denaturation
KW - Recombinant protein
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U2 - 10.1016/j.bej.2013.06.004
DO - 10.1016/j.bej.2013.06.004
M3 - Article
AN - SCOPUS:84880101543
VL - 77
SP - 177
EP - 182
JO - Biochemical Engineering Journal
JF - Biochemical Engineering Journal
SN - 1369-703X
ER -