|
| Parameter | Value |
|
| General assumptions for the facility |
| rBuChE production level | 25 kg rBuChE/year (bulk) |
| rBuChE doses per year | 62,500 doses/year at 400 mg/dose |
| Downstream Recovery/purification yield | 20% |
| Annual operating days | 330 days |
| rBuChE production in plants following infiltration/incubation | 125 kg rBuChE/year |
| Batch cycle time (time between start of new infiltration batches) | 7 days |
| Batches per year | 47 |
| Base case rBuChE expression level | 500 mg/kg FW at ~7 days after infiltration |
| Facility lifetime | 15 years |
| Depreciation | Straight line over 10 years, 5% direct fixed capital salvage |
| Working capital | 30 days of labor, materials, utilities, waste treatment |
| Lab/QC/QA costs | 2% of total labor costs for plant growth and agroinfiltration sections, 15% of total labor costs for recovery and purification section |
| Start-up/validation costs | 5% direct fixed capital |
|
| Assumptions for indoor plant growth section |
| Mass per plant at 5 weeks | 0.02 kg FW/plant |
| Tray size | 4 ft × 4 ft |
| Number of plants per tray | 256 |
| Tobacco seed cost | $0.001/seed |
| Age at infiltration | 4 weeks |
| Total number of plant batches in inventory | 5 batches (just seeded, 1 wk old, 2 wk old, 3 wk old, 4 wk old—ready to infiltrate, 5 wk old—infiltrated, incubated and ready to harvest) |
| Total number of plants in inventory | ~1.3 million |
| Total plant growth area | 83,320 ft2 total, or 10 levels with 8,332 ft2 footprint |
| LED fixture costs | $40/ft2 plant growth area (including capital cost factors for plant growth area as shown in Table 2) |
| LED energy costs |
20 W/ft2 |
|
| Assumptions for Agrobacterium production, vacuum infiltration, and plant incubation section |
| Agro “loading”—mass of recombinant Agrobacterium to mass of plant tissue | 0.00001 kg dry weight (dw) bacteria/kg FW plant biomass |
| Agrobacterium biomass density at 12 hours of culture | 2.6 g dw/L = 0.0026 kg dw/L |
| Inoculum density used in seed train | 1% V/V |
| Dilution factor between agroinfiltration solution and agrobacterial production fermentor | 78 |
| Percent weight change in plant tissue following vacuum agroinfiltration | 30% |
| Trays processed per vacuum chamber (30 ft) per day | 336 trays/chamber/day |
| “Excess” Agrobacterium solution used | 87% of total infiltration solution |
| Incubation time for infiltrated plants | ~7 days |
|
| Assumptions for the rBuChE recovery and purification section |
| Overall yield in downstream processes | 20% |
| Harvesting rate | 3 trays/minute |
| IEX chromatography | |
| Binding capacity | 20 mg/mL |
| Resin cost | $1,839/L |
| Number of reuse cycles | 100 |
| Affinity chromatography | |
| Binding capacity | 3 mg/mL |
| Resin cost | $10,000/L |
| Number of reuse cycles | 30 |
|
