Device for Converting Elastin-Like Polypeptide Aggregate to Soluble Form

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Device for Converting Elastin-Like Polypeptide
Aggregate to Soluble Form

• Eric Lee, John Harrison,
• Albert Kwansa, Sasha Cai Lesher-Perez
• Client Dr. Darin Furgeson
• Advising Professor Dr. William Murphy

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Overview of Presentation

• Client research
• Elastin-like polypeptides (ELP) introduction
• Project design statement
• Design specifications and assumptions
• Proposed designs
• Future Work
• Answer questions

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Client Information

• Dr. Darin Furgeson from Pharmaceutical Sciences
  Department
• Novel cancer treatment delivery system with ELP
• Target specific cells
• Non-viral
• Lower toxicity

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Background Information on ELP

• Synthetic protein
• Repeating pentapeptide sequence
• (Val-Pro-Gly-Xaa-Gly)
• Hydrophobic interaction
• Thermal Responsive (transition temperature, Tt)
• Non-viral vector for cancer treatment delivery

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Current Extraction Process

• Transformation of plasmid into E. coli
• Transcription and translation of ELP
• Extraction of ELP from inclusion body
• (differential centrifugation)
• ELP aggregation/separation from soluble DNA
• Re-suspension

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Project Design Statement

• Design a device with temperature control, salt
  extraction, and particle reduction capabilities
  to enhance solubility of ELP aggregate while
  minimizing product loss.

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Design Specifications

• 75-80 yield of ELP
• Maintenance below Tt
• Durable material selection
• Reduction of particle size
• Portability

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Assumptions for Design

• Mechanical stress
• Various agitation methods have little
  degradative effects on ELP structure.

• Heat Fluctuations
• Changes to extreme
• temperatures will not
• damage ELP structural integrity.

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Design 1
Device components
Operation
Cone-shaped drill

1. Freeze ELP in funnel
2. Initiate cone-shaped drill
3. Fall through non-stick tube
4. Descend into reservoir of cold PBS

Funnel
Temperature control tank
Chilled PBS reservoir with inlet
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Design 1

• Pros
• Frozen ELP more manageable
• Cons
• Electric power requirement due to use of motor.
• Multiple parts requiring cooling.

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Design 2

• Operation
• Place Frozen ELP on spinning blade
• Force ELP through with plunger
• Shavings fall in churning PBS bath

Blade
PBS Churner
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Design 2

• Pros
• Requires less force to break down frozen ELP
• Less manual intensive
• Faster, cleaner
• Cons
• Very dynamic, many moving parts
• Costly

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Design 3

• Operation
• Coat screen with ELP aggregate
• Submerge in cold PBS
• Rotate at high speed/high torque

Motor
Shaft
PBS Tub
Non-stick Screen
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Design 3

• Pros
• No frozen ELP
• Simple, easy to operate
• Cons
• Material must meet unique design specifications
• Force required to push ELP out may be much
  greater than anticipated

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Future Studies

• Analyze/test feasibility of all three designs
• Determine precise design specifications
• Determine cost of manufacturing each design
• Analyze complexity of each design
• Refine designs for prototype

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• Questions?

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More Slides

• Chemotherapy
• Non-specific
• Highly toxic
• Patients lose hair, feels sick, have no energy
• ELP delivery system
• ELP tagged medicine
• Heat pad
• Endocytosis
• Lysosome