Immobilization of transgenic plant cells towards bioprinting for production of a recombinant biodefense agent. Issue 10 (15th August 2021)

• Record Type: Journal Article
• Authors: Varma, Anika; Gemeda, Hawi B.; McNulty, Matthew J.; McDonald, Karen A.; Nandi, Somen; Knipe, Jennifer M.
• Is Part Of: Biotechnology journal. Volume 16:Issue 10(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-08-15
• Subjects: bioprinting -- butyrylcholinesterase -- cell immobilization -- continuous bioproduction -- hydrogel -- transgenic rice cell culture; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.202100133
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract: Transgenic rice cells ( Oryza sativa ) producing recombinant butyrylcholinesterase (BChE) as a prophylactic/therapeutic against organophosphate nerve agent poisoning, cocaine toxicity, and neurodegenerative diseases like Alzheimer's were immobilized in a polyethylene glycol‐based hydrogel. The cells were sustained for 14 days in the semi‐solid matrix, undergoing a growth phase from days 0–6, a BChE production phase in sugar‐free medium from days 6–12, and a growth/recovery phase from days 12–14. Throughout this period, the cells maintained similar viability to those in suspension cultures and displayed analogous sugar consumption trends. The rice cells in the hydrogel also produced a significant amount of active BChE, comparable to the levels produced in liquid cultures. A considerable fraction of this BChE was secreted into the media, allowing for easier product separation. To the best of our knowledge, this proof‐of‐concept is the first report of immobilization of recombinant plant cells for continuous production of high‐value heterologous proteins. This work serves as a foundation for further investigation towards plant cell bioprinting and the development of a simple, efficient, robust, modular, and potentially field‐deployable bioreactor system for the manufacture of biologics. Graphical Abstract and Lay Summary: Transgenic rice cells were combined with a polyethylene glycol tetra‐acrylate (PEGTA) and lithium phenyl‐2, 4, 6‐trimethylbenzoylphosphinate (LAP) bioink and cured with UV light to construct an immobilized cell‐based protein production system. The cells were maintained for 14 days in the hydrogel matrix and were induced to actively make and secrete recombinant butyrylcholinesterase, a complex enzyme that irreversibly binds to and can hydrolyze organophosphate. This proof‐of‐concept study showcases the use of immobilized and potentially bioprintable plant cells to produce high‐value proteins with prophylactic and therapeutic applications.