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