Team:Cornell/Future

From 2011.igem.org

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Our next goal would be to further reduce the cost of manufacturing this cell-free system.  To keep most of our current system's benefits while increasing cost-efficiency, we'd like to describe a novel system involving micro-carriers.
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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-efficiency, we'd like to describe a novel system involving micro-carriers.
====Main System Changes:====
====Main System Changes:====
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====Description of changes====
====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.
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.
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Mesh filters between vats, will prevent these microcarrier beads from traveling across vats. The substrate will sequentially undergo specific reactions in each vat until the final product is obtained.  One of the largest problems in this solution is mixing the beads with the 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 would have been in contact with enzymes for a long time mostly reacted.  The solution at the top of the vat will then be fed to the bottom of the next vat.
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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:====
====This system will retain benefits of our existing project:====
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# Minimum side reactions.
# Minimum side reactions.
# Cell-free.
# Cell-free.
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# Modular reactions.
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# Continuous, yet modular reactions.
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# Continuous, yet modular reactions
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====Additional benefits:====
====Additional benefits:====
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====Potential drawbacks:====
====Potential drawbacks:====
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# Decrease in solution mixture homogeneity due to larger reaction volumes.
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# Decrease in reaction mixture homogeneity due to larger reaction volumes.
# Less control over reaction conditions.
# Less control over reaction conditions.
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Revision as of 02:02, 29 September 2011

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-efficiency, we'd like to describe a novel system involving micro-carriers.

Main System Changes:

  1. Reaction Chamber: Replace microfluidic chips with large vats
  2. 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:

  1. Lower purification costs.
  2. Minimum side reactions.
  3. Cell-free.
  4. Continuous, yet modular reactions.

Additional benefits:

  1. Cheaper to scale up manufacturing process.
  2. Larger reaction volumes.

Potential drawbacks:

  1. Decrease in reaction mixture homogeneity due to larger reaction volumes.
  2. Less control over reaction conditions.

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