1. 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

2. Overview of Presentation  Client research  Elastin-like polypeptides (ELP) introduction  Project design statement  Design specifications and assumptions  Proposed designs  Future Work  Answer questions

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

4. 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

5. 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

6. 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.

7. Design Specifications  75-80% yield of ELP  Maintenance below Tt  Durable material selection  Reduction of particle size  Portability

8. 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.

9. Design #1 Cone-shaped drill Funnel Temperature control tank Chilled PBS reservoir with inlet Device components 1. Freeze ELP in funnel 2. Initiate cone-shaped drill 3. Fall through non-stick tube 4. Descend into reservoir of cold PBS Operation

10. Design #1 Pros: Frozen ELP: more manageable Cons: Electric power requirement due to use of motor. Multiple parts requiring cooling.

11. Design #2  Operation Place Frozen ELP on spinning blade Force ELP through with plunger Shavings fall in churning PBS bath Blade PBS Churner

12. Design #2  Pros Requires less force to break down frozen ELP Less manual intensive Faster, cleaner  Cons Very dynamic, many moving parts Costly

13. Design #3  Operation Coat screen with ELP aggregate Submerge in cold PBS Rotate at high speed/high torque Non-stick Screen PBS Tub Motor Shaft

14. 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

15. 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

16. Questions?

17. 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