ELASTINELASTINERIC LEEERIC LEE
-LIKE PO-LIKE POALBERT KWANSAALBERT KWANSA
ADVISOR: PROFESSORADVISOR: PROFESSOR
LYPEPTILYPEPTIJOHN HARRISONJOHN HARRISON
WILLIAM MURPHYWILLIAM MURPHY
DE RE-DE RE- SASHA CAI LESHER –SASHA CAI LESHER –
CLIENT: DR. DARIN FU CLIENT: DR. DARIN FU
SOLUBILSOLUBILPPÉREZÉREZ
RGESONRGESON
ISATIONISATION
AbstractAbstract
The purpose of our final design is to replace the The purpose of our final design is to replace the current manually intensive ELP solubilisation method current manually intensive ELP solubilisation method with a faster, more efficient method that will have with a faster, more efficient method that will have higher product recovery percentage. We came up with higher product recovery percentage. We came up with three design ideas, and after research and design three design ideas, and after research and design considerations, we incorporated aspects from all three considerations, we incorporated aspects from all three proposed design in our final prototype. The final proposed design in our final prototype. The final prototype that we constructed meets design prototype that we constructed meets design specifications, but there is future work that could be specifications, but there is future work that could be done to improve the effectiveness of the device. done to improve the effectiveness of the device.
Project MotivationProject Motivation
ChemotherapyChemotherapyHighly ToxicHighly ToxicNon-SpecificNon-Specific
Elastin-Like PolypeptideElastin-Like PolypeptideCell SpecificCell SpecificNon-ViralNon-ViralLower ToxicityLower Toxicity
ELP PropertiesELP Properties Synthetic ProteinSynthetic Protein
Repeating sequence of five Repeating sequence of five principle amino acids (Val-Pro-principle amino acids (Val-Pro-Gly-Xaa-Gly)Gly-Xaa-Gly)
Responds to Responds to temperaturetemperature Transition temperature (Tt)Transition temperature (Tt)
Hydrophobic Interaction Hydrophobic Interaction aggregationaggregation
H3C
CH3
NH2
O
OH
HN
HO
O
NH2HO
O
Valine
Proline
Glycine
Current MethodsCurrent Methods
Extraction of Cell from E. coliExtraction of Cell from E. coli
Purification with addition of SaltPurification with addition of Salt
Re-suspensionRe-suspension Require 12 hours of manual labor to Require 12 hours of manual labor to
re-suspend 300~600mgre-suspend 300~600mg
Project Design StatementProject Design Statement
Design a device with temperature Design a device with temperature control, salt extraction, and particle control, salt extraction, and particle reduction capabilities to enhance solubility reduction capabilities to enhance solubility of ELP aggregate while minimizing product of ELP aggregate while minimizing product loss.loss.
Project Operation GoalProject Operation Goal
75-80% yield of ELP after 75-80% yield of ELP after solubilisationsolubilisation
Maintain temperature below TtMaintain temperature below Tt
Durable material selectionDurable material selection
Reduction of particle sizeReduction of particle size
AutomationAutomation
Design GridDesign GridPrototypePrototype CostCost
(1-10)(1-10)
ProductProduct
RecoveryRecovery
(1-10)(1-10)
FeasibilityFeasibility
(1-10)(1-10)
EaseEase
OfOf
OperationOperation
(1-10)(1-10)
DurabilityDurability
(1-5)(1-5)
OverallOverall
99 77 88 88 55 3737
33 66 44 44 55 2222
22 66 44 55 55 2222
Final DesignFinal 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
1.45 cm
2.5 cm
1.54 cm
Dimensions:
18 cm
Viscosity of honey = 15.0 NViscosity of honey = 15.0 N··s/ms/m2 2 (van den Berg, Arie)(van den Berg, Arie)
= 515 rad/s ; r= 515 rad/s ; rmaxmax = 0.0075 m ; Thickness = 0.0075 m = 0.0075 m ; Thickness = 0.0075 m
Area = 0.001005 mArea = 0.001005 m22
StressStressmaxmax = Viscosity*(r = Viscosity*(rmaxmax**/Thickness/Thickness
= 7728 N/m= 7728 N/m22
ForceForcemaxmax = Stress* Area = Stress* Area
= 3.88 N= 3.88 N
Final Design Calculations Final Design Calculations
Fmax
Fmax
Thickness
rmax
Final DesignFinal DesignProsPros
Can fit within a test tubeCan fit within a test tube
Minimal loss of ELPMinimal loss of ELP
Integrated power switchIntegrated power switch
Interchangeable head- pieceInterchangeable head- piece
ConsCons
Requires continuous Requires continuous
battery replacementbattery replacement
Currently, smaller test Currently, smaller test
tube sizes cannot be tube sizes cannot be
accommodatedaccommodated
Cut out paddle from aluminum sheets Cut out paddle from aluminum sheets (0.025”)(0.025”)
Machined steel rod to 75mm ( Machined steel rod to 75mm ( ΦΦ = 0.081”) = 0.081”)
Drilled and slotted an aluminum adapter to Drilled and slotted an aluminum adapter to connect rod, motor, and paddleconnect rod, motor, and paddle
Bore out septa rubber cap to affix device to Bore out septa rubber cap to affix device to 15 ml test tube 15 ml test tube
Prototype Prototype ManufacturingManufacturing
TestingTesting Viscosity of honey and Viscosity of honey and
viscous ELP comparableviscous ELP comparable
Verified torsion capability in Verified torsion capability in viscous materialviscous material
Tested in aggregated ELP Tested in aggregated ELP substitute (rubber shavings) substitute (rubber shavings) observed interaction with observed interaction with waterwater
Future ModificationsFuture Modifications Variety of head piecesVariety of head pieces
Different shapesDifferent shapes Varying sizesVarying sizes Drill-like heads Drill-like heads Splash guardSplash guard
Non-stick componentsNon-stick components TeflonTeflon SigmaCoteSigmaCote
AcknowledgementsAcknowledgements
We would like to thank our advisor, Professor William We would like to thank our advisor, Professor William
Murphy, for his guidance and encouragement during the Murphy, for his guidance and encouragement during the
semester and we would like to express our gratitude towards semester and we would like to express our gratitude towards
our client, Dr. Darin Furgeson, for his support, laboratory our client, Dr. Darin Furgeson, for his support, laboratory
resources, and for giving us the opportunity to work on a resources, and for giving us the opportunity to work on a
project that could ultimately contribute to medical treatment.project that could ultimately contribute to medical treatment.
ReferencesReferences Urry, Dan W. Physical Chemistry of Biological Free Energy Urry, Dan W. Physical Chemistry of Biological Free Energy
Transduction as Demonstrated by Elastic Protein-Based Transduction as Demonstrated by Elastic Protein-Based Polymers, Polymers, Journal of Physical ChemistryJournal of Physical Chemistry 1997 1997
Meyer, D., Trabbic-Carlson K., and Chilkoti A.,Protein Purification Meyer, D., Trabbic-Carlson K., and Chilkoti A.,Protein Purification by Fusion with an Environmentally Responsive Elastin-Like by Fusion with an Environmentally Responsive Elastin-Like Polypeptide: Effect of Polypeptide Length on the Purification Polypeptide: Effect of Polypeptide Length on the Purification of Thioredoxi, of Thioredoxi, Biotechnology Biotechnology 20012001
Meyer, D. and Chilkoti, A., Purification of recombinant proteins by Meyer, D. and Chilkoti, A., Purification of recombinant proteins by fusion with thermally responsive polypeptides, fusion with thermally responsive polypeptides, NatureNature 1999 1999
van den Berg, Arie. van den Berg, Arie. The production of "good" creamed honeyThe production of "good" creamed honey. . Retrieved December 1, 2005, from The University of Retrieved December 1, 2005, from The University of Queensland, Department of Chemical Engineering Web site: Queensland, Department of Chemical Engineering Web site: http://www.cheque.uq.edu.au/ugrad/theses/1998/pdf/ARIE.pdf http://www.cheque.uq.edu.au/ugrad/theses/1998/pdf/ARIE.pdf