Transcript Project Motivation

ELASTIN
ERIC LEE
-LIKE PO
ALBERT KWANSA
ADVISOR: PROFESSOR
LYPEPTI
JOHN HARRISON
WILLIAM MURPHY
DE RESASHA CAI LESHER –
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-ProGly-Xaa-Gly)
CH3 O
Valine
H3 C
OH
NH2
HO
 Responds to temperature
Proline
O
 Transition temperature (Tt)
 Hydrophobic Interaction 
aggregation
Glycine
HN
HO
NH2
O
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
Prototype Cost Product Feasibility Ease
Durability Overall
(1-10) Recovery (1-10)
Of
(1-5)
(1-10)
Operation
(1-10)
9
7
8
8
5
37
3
6
4
4
5
22
2
6
4
5
5
22
Final Design
Dimensions:
Device Components:
Mabuchi Motor
(FF-130SH)
Rubber Top
Steel
Shaft
Aluminum
Paddle
(Mixer)
Power Supply
(9 volt battery)
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
Fmax
rmax
Forcemax = Stress* Area
= 3.88 N
w
Thickness
Fmax
Final Design
Pros
Cons
 Can fit within a test tube
 Requires continuous
 Minimal loss of ELP
battery replacement
 Integrated power switch
 Currently, smaller test
 Interchangeable head-
tube sizes cannot be
piece
accommodated
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
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:
http://www.cheque.uq.edu.au/ugrad/theses/1998/pdf/ARIE.pdf