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ELASTIN ERIC LEE

ELASTIN ERIC LEE. -LIKE PO ALBERT KWANSA ADVISOR: PROFESSOR. LYPEPTI JOHN HARRISON WILLIAM MURPHY. DE RE- SASHA CAI LESHER – CLIENT: DR. DARIN FU. SOLUBIL P ÉREZ RGESON. ISATION. Abstract.

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ELASTIN ERIC LEE

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  1. ELASTIN ERIC LEE

  2. -LIKE PO ALBERT KWANSA ADVISOR: PROFESSOR

  3. LYPEPTI JOHN HARRISON WILLIAM MURPHY

  4. DE RE- SASHA CAI LESHER – CLIENT: DR. DARIN FU

  5. SOLUBIL PÉREZ RGESON

  6. ISATION

  7. Abstract The purpose of our final design is to replace the current manually intensive ELP solubilisation method with a faster, more efficient method that will have higher product recovery percentage. We came up with three design ideas, and after research and design considerations, we incorporated aspects from all three proposed design in our final prototype. The final prototype that we constructed meets design specifications, but there is future work that could be done to improve the effectiveness of the device.

  8. Project Motivation Chemotherapy Highly Toxic Non-Specific Elastin-Like Polypeptide Cell Specific Non-Viral Lower Toxicity

  9. ELP Properties • Synthetic Protein • Repeating sequence of five principle amino acids (Val-Pro-Gly-Xaa-Gly) • Responds to temperature • Transition temperature (Tt) • Hydrophobic Interaction  aggregation

  10. Current Methods • Extraction of Cell from E. coli • Purification with addition of Salt • Re-suspension • Require 12 hours of manual labor to re-suspend 300~600mg

  11. 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.

  12. Project Operation Goal • 75-80% yield of ELP after solubilisation • Maintain temperature below Tt • Durable material selection • Reduction of particle size • Automation

  13. Design Grid

  14. Final Design Dimensions: Device Components: 1.54 cm Mabuchi Motor (FF-130SH) 1.45 cm 2.5 cm Rubber Top 8.0 cm Steel Shaft 18 cm 1.45 cm Power Supply (9 volt battery) Aluminum Paddle (Mixer) 6.7 cm 1.5 cm

  15. Final Design Calculations Viscosity of honey = 15.0 N·s/m2 (van den Berg, Arie) w = 515 rad/s ; rmax = 0.0075 m ; Thickness = 0.0075 m Area = 0.001005 m2 Stressmax = Viscosity*(rmax*w)/Thickness = 7728 N/m2 Forcemax = Stress* Area = 3.88 N Fmax rmax w Thickness Fmax

  16. Pros Can fit within a test tube Minimal loss of ELP Integrated power switch Interchangeable head- piece Cons Requires continuous battery replacement Currently, smaller test tube sizes cannot be accommodated Final Design

  17. Prototype Manufacturing Cut out paddle from aluminum sheets (0.025”) Machined steel rod to 75mm ( Φ = 0.081”) Drilled and slotted an aluminum adapter to connect rod, motor, and paddle Bore out septa rubber cap to affix device to 15 ml test tube

  18. Testing • Viscosity of honey and viscous ELP comparable • Verified torsion capability in viscous material • Tested in aggregated ELP substitute (rubber shavings) observed interaction with water

  19. Future Modifications • Variety of head pieces • Different shapes • Varying sizes • Drill-like heads • Splash guard • Non-stick components • Teflon • SigmaCote

  20. Acknowledgements We would like to thank our advisor, Professor William Murphy, for his guidance and encouragement during the semester and we would like to express our gratitude towards our client, Dr. Darin Furgeson, for his support, laboratory resources, and for giving us the opportunity to work on a project that could ultimately contribute to medical treatment.

  21. References • Urry, Dan W. Physical Chemistry of Biological Free Energy Transduction as Demonstrated by Elastic Protein-Based Polymers, Journal of Physical Chemistry 1997 • Meyer, D., Trabbic-Carlson K., and Chilkoti A.,Protein Purification by Fusion with an Environmentally Responsive Elastin-Like Polypeptide: Effect of Polypeptide Length on the Purification of Thioredoxi, Biotechnology 2001 • Meyer, D. and Chilkoti, A., Purification of recombinant proteins by fusion with thermally responsive polypeptides, Nature 1999 • van den Berg, Arie. The production of "good" creamed honey. Retrieved December 1, 2005, from The University of Queensland, Department of Chemical Engineering Web site: http://www.cheque.uq.edu.au/ugrad/theses/1998/pdf/ARIE.pdf

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