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Team:Cornell/Future
From 2011.igem.org
Project Description |
Future Directions |
Business Development |
Outreach/HP |
Safety
Our next goal would be to further reduce the cost of manufacturing in this cell-free system. To keep most of our current system's benefits while increasing cost-effectiveness, we'd like to describe a novel system involving micro-carriers.
Main System Changes:
- Reaction Chamber: Replace microfluidic chips with large vats
- Enzyme-coated surface: Replace streptavidin-coated microfluidic channels with streptavidin-coated PDMS beads
Description of changes:
PDMS beads coated with our avi-tagged enzymes would be placed in large vats. Each vat would contain beads coated with a different enzyme and these vats would be arranged in series according to our substrate's biosynthetic pathway. Mesh filters between vats will prevent these microcarrier beads from traveling across vats. The substrate will sequentially undergo enzyme-mediated reactions in each vat until the final product is obtained. One of the largest challenges for this system is thoroughly mixing the PDMS beads with reactants. One possible solution is to flow substrate through the bottom of the vat, then slowly mix it while pushing the solution up using a spiraling turbine. Once the solution reaches the top, the substrates will have been in contact with enzymes for a long time, maximizing opportunity for the reaction to occur. The solution at the top of the vat will then be fed to the bottom of the next vat.
This system will retain benefits of our existing project:
- Lower purification costs.
- Minimum side reactions.
- Cell-free.
- Continuous, yet modular reactions.
Additional benefits:
- Cheaper to scale up manufacturing process.
- Larger reaction volumes.
Potential drawbacks:
- Decrease in reaction mixture homogeneity due to larger reaction volumes.
- Less control over reaction conditions.
