Semicontinuous bioreactor production of recombinant butyrylcholinesterase in transgenic rice cell suspension cultures
2016
Jasmine Marie Corbin | Bryce Isao Hashimoto | Kalimuthu eKaruppanan | Zachary Ryan Kyser | Liying eWu | Brian A Roberts | Amy R Noe | Raymond L Rodriguez | Karen Ann McDonald | Somen eNandi
An active and tetrameric form of recombinant butyrylcholinesterase (BChE), a large and complex human enzyme, was produced via semicontinuous operation in a transgenic rice cell suspension culture. After transformation of rice callus and screening of transformants, the cultures were scaled up from culture flask to a lab scale bioreactor. The bioreactor was operated through two phases each of growth and expression. The cells were able to produce BChE during both expression phases, with a maximum yield of 1.6 mg BChE/L of culture during the second expression phase. Cells successfully regrew during a 5-day growth phase. A combination of activity assays and Western blot analysis indicated production of an active and fully assembled tetramer of BChE.
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