ELASTIN ERIC LEE

-LIKE PO ALBERT KWANSA ADVISOR: PROFESSOR

LYPEPTI JOHN HARRISON WILLIAM MURPHY

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

SOLUBIL PÉREZ RGESON

ISATION

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.

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

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

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

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.

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

Design Grid PrototypeCost(1-10)ProductRecovery(1-10)Feasibility(1-10)EaseOfOperation(1-10)Durability(1-5)Overall

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

Viscosity of honey = 15.0 N·s/m 2 (van den Berg, Arie)  = 515 rad/s ; r max = m ; Thickness = m Area = m 2 Stress max = Viscosity*(r max *  /Thickness = 7728 N/m 2 = 7728 N/m 2 Force max = Stress* Area = 3.88 N = 3.88 N Final Design Calculations F max  Thickness r max

Final Design 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

 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 Prototype Manufacturing

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

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

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.

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: