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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. Overview of Presentation. Client research
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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
Overview of Presentation • Client research • Elastin-like polypeptides (ELP) introduction • Project design statement • Design specifications and assumptions • Proposed designs • Future Work • Answer questions
Client Information • Dr. Darin Furgeson from Pharmaceutical Sciences Department • Novel cancer treatment delivery system with ELP • Target specific cells • Non-viral • Lower toxicity
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
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
Design a device with temperature control, salt extraction, and particle reduction capabilities to enhance solubility of ELP aggregate while minimizing product loss. Project Design Statement
Design Specifications • 75-80% yield of ELP • Maintenance below Tt • Durable material selection • Reduction of particle size • Portability
Mechanical stress Various agitation methods have little degradative effects on ELP structure. Heat Fluctuations Changes to extreme temperatures will not damage ELP structural integrity. Assumptions for Design
Freeze ELP in funnel Initiate cone-shaped drill Fall through non-stick tube Descend into reservoir of cold PBS Design #1 Device components Operation Cone-shaped drill Funnel Temperature control tank Chilled PBS reservoir with inlet
Design #1 • Pros: • Frozen ELP: more manageable • Cons: • Electric power requirement due to use of motor. • Multiple parts requiring cooling.
Design #2 • Operation • Place Frozen ELP on spinning blade • Force ELP through with plunger • Shavings fall in churning PBS bath Blade PBS Churner
Design #2 • Pros • Requires less force to break down frozen ELP • Less manual intensive • Faster, cleaner • Cons • Very dynamic, many moving parts • Costly
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
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
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
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