2. Agronomy Team:
Don C. Wilkerson
Isaac Wong
Stephanie Akins

3. Nicotiana benthamiana
Nicotiana: A genus of short-leafed plants
in the nightshade family including
Nicotiana tabacum
Nicotiana benthamiana: A close relative
of tobacco belonging to the same genus
of short-leafed plants and used as a
model organism
Nicotiana tabacum: The common tobacco
plant cultivated worldwide to produce
tobacco leaf for cigarettes

4. Grow Pods:

5. Production Protocol:
Nicotiana bentamiama plants are grown using a Standard Production Protocol
(SPP) developed by GCON/Caliber Biotheraputics. This SPP has evolved
through a series of platform optimization studies/trials/evaluations, designed
to reach targeted expression levels for a broad range of monoclonal
antibodies.

6. Seeding/Plug Production
Transplant 21 Days
Grow 10 Days
Infiltration
Post Infiltration 7 Days
Harvest
Standard Production Protocol
Total Production Time = 38 Days

7. Seeding:
The SPP begins by direct seeding in to Styrofoam
plug trays, each containing 240 rockwool plugs.
Prior to seeding, plugs are pre-soaked with a pH
adjusted nutrient solution to ensure optimum
germination and growth. Following seeding, trays
are placed in a controlled environment Pod at
25⁰C and 75% humidity. Supplemental light is
provided 24 hours/day by LED fixtures, delivering
60 microMoles/second/m2 at tray height. All
trays are sub-irrigated 3X/day at 8 hour
intervals, using a pH adjusted, hydroponic
nutrient solution.
90% of all seed germinate within 3 – 5 days.
Seedlings (plugs) will remain in this environment
for approximately 21 days.

8. 21 day old plugs are then hand
transplanted in to trays containing pre-
dibbled sheets of an expanded
polypropylene growing substrate
(SuretoGrow). Each tray is fitted with a
perforated lid, designed to hold the
substrate/plants in place during vacuum
infiltration. Each Grow tray contains 60
plants.
Transplanting:

9. Transplanted trays are then placed in a
controlled environment Pod at 25⁰C and
60% humidity. Supplemental light is
provided 24 hours/day by LED
fixtures, delivering 120
microMoles/second/m2 at tray height.
All trays are irrigated 3X/day at 8 hour
intervals, using a pH
adjusted, hydroponic nutrient solution.
Plants are grown under these conditions
for 10 days prior to vacuum infiltration.
Grow:

10. Infiltration:

11. Following vacuum infiltration, plants are placed
in a post-infiltration (PI) Pod at 25⁰C and 60%
humidity. Supplemental light is provided 18
hours/day by LED fixtures, delivering 120
microMoles/second/m2 at tray height. All trays
are irrigated 3X/day at 8 hour intervals, using a
pH adjusted, hydroponic nutrient solution.
Plants are grown under these conditions for 10
days prior to harvest.--
Post Infiltration:

12. Harvest: