Proteomic Profiling of Interplay Between Agrobacterium tumefaciens and Nicotiana benthamiana for Improved Molecular Pharming Outcomes

Prudhomme, Nicholas; Krieger, Jonathan R.; McLean, Michael D.; Cossar, D.; Geddes‐McAlister, Jennifer

doi:10.1007/978-1-0716-2124-0_19

Cited by 1 publication

(1 citation statement)

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“…Protein extractions were performed as previously described (Prudhomme et al ., 2022). Briefly, frozen leaf tissue (30 mg) was suspended in 100 m m Tris–HCl (pH 8.5) containing one protease inhibitor cocktail tablet (Roche, Basel, Switzerland).…”

Section: Methodsmentioning

confidence: 99%

Bacterial growth‐mediated systems remodelling of Nicotiana benthamiana defines unique signatures of target protein production in molecular pharming

Prudhomme,

Pastora,

Thomson

et al. 2024

Plant Biotechnology Journal

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SummaryThe need for therapeutics to treat a plethora of medical conditions and diseases is on the rise and the demand for alternative approaches to mammalian‐based production systems is increasing. Plant‐based strategies provide a safe and effective alternative to produce biological drugs but have yet to enter mainstream manufacturing at a competitive level. Limitations associated with batch consistency and target protein production levels are present; however, strategies to overcome these challenges are underway. In this study, we apply state‐of‐the‐art mass spectrometry‐based proteomics to define proteome remodelling of the plant following agroinfiltration with bacteria grown under shake flask or bioreactor conditions. We observed distinct signatures of bacterial protein production corresponding to the different growth conditions that directly influence the plant defence responses and target protein production on a temporal axis. Our integration of proteomic profiling with small molecule detection and quantification reveals the fluctuation of secondary metabolite production over time to provide new insight into the complexities of dual system modulation in molecular pharming. Our findings suggest that bioreactor bacterial growth may promote evasion of early plant defence responses towards Agrobacterium tumefaciens (updated nomenclature to Rhizobium radiobacter). Furthermore, we uncover and explore specific targets for genetic manipulation to suppress host defences and increase recombinant protein production in molecular pharming.

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Section: Methodsmentioning

confidence: 99%