Office for Technology Licensing and Industry Collaboration ● Tufts School of Engineering Polymer...

Office for Technology Licensing and Industry Collaboration ● http://techtransfer.tufts.edu

Tufts School of EngineeringTufts School of Engineering

Polymer Discovery Via Polymer Discovery Via Microfluidic Enzymatic SynthesisMicrofluidic Enzymatic Synthesis

Prof. Peter Y. WongProf. David KaplanTufts University - Medford, MA

October 3, 2006


OverviewOverview

• Markets• Needs• Problems• Solution• Team• Next Steps• Summary

Biochemical Science

Enzyme Polymer science 1970-

Synthesis 1990-

Micro-fabrication

Microsystems Technology

Engineering

Mechanical Electronics


Many MarketsMany Markets

• Any market that benefits from new biochemicals• Improved Foods

– Additives, Modification, Nutrition

• Green Chemistry– Agricultural, Packaging, Analysis

• New Medicines– Topical, Digested, Structural


Markets Needs vs.WantsMarkets Needs vs.Wants

• New products– Better– Faster– Cheaper– Differentiated

• Macro and Micromolecules needed– New material– New processes

• Focus on new polymers and processes


Problems and RisksProblems and Risks

• Current Polymer Discovery Process– Long time with process and people– High costs and large resources needed– FDA, EPA stringent regulations– Limited research to commercialization

• Alternate Approaches– Nanoscience/technologies - far in future?– Biomimetics/Bioinspiration - hit or miss?– Microengineering/fluidics - scalability?


Our SolutionOur Solution

• Achieve “Green ( ) Polymer Chemistry” through – Enzymatic Synthesis and– Microfluidics

• Enzymatic polymerizations can produce products– via mild reaction conditions w/o toxic reagents– in an environmentally friendly synthetic process– that can be scaled from microscale to macroscale

• Target macromolecules include– polysaccharides, polyesters, polycarbonates, poly(amino acid)s,

polyaromatics, and/or vinyl polymers.


Microfluidic Enzymatic CascadeMicrofluidic Enzymatic Cascade

• Universal Lab-On-Chip is very far away• Application Specific Integrated Microfluidic

(ASIM) device • Example ASIM –

– produce vitamin C enriched polymers (PMMA) polymer – has both scientific and market value.

Antioxidant Polymer

Natural antioxidant

Monomer

Enzyme 1

Enzyme 2


PMMA PolymerPMMA Polymer

• Disruptive Technology in Packaging– Vitamin C enriched polymers can replace butylated hydroxy

anisole (BRA) and butylated hydroxy toluene (BHT) - FDA limits conc. To 0.02%.

• New Topical Medicine – Antioxidants are considered important in reducing aging-related

phenomena by providing protection against free radicals.

• Nutraceutical Supplementation– Ascorbic acid may have an overall positive impact on public

health because humans lack the ability to synthesize vitamin C


ASIMASIM

– Goals:

– Two enzymatic cascade reactions to produce PMMA

– low cost devices made of poly(dimethylsiloxane) (PDMS)

– efficient method to optimize process with external controls


Translation from Abstract to HardwareTranslation from Abstract to Hardware

Reaction Vessel #1

React with lipase

Reaction Vessel #2

React with HRP

Input:

monomer

in solvent

Input:

ascorbic acid

Input : hydrogen

peroxide

Output: unreacted

ascorbic acid

Output: unreacted

hydrogen peroxide

OutputReaction Vessel #1Input:

in solvent

Input: ascorbic acid

Input

: hydrogen peroxide

Output: unreacted

ascorbic acid

Output: unreacted

hydrogen peroxide

Output

polymer

::

ascorbic acid

lipase

HRP

monomer

AA-Monomer

P-AA-MMA

Check Valve#1 #2

AA- Ascorbic Acid

MMA- Methyl Methacrylate

PMMA- Poly (Methyl Methacrylate)

P-AA-MMA – Ploy L-Ascorbic Methyl Methacrylate

HRP – Horse Radish Peroxidase

monomer

Antioxidant polymer

ascorbic acid

lipase

AA-Monomer

Hydrogen peroxide

HRP

AA-Monomer


Improved VersionImproved Version

Vessel 2 Poly L-Ascorbyl Methyl methacrylate(P-AA-MMA) (G.2)

Vessel 1 L-Ascrbyl Methyl methacrylate(AA-MMA) (G 1)

1.77ul2,4-pentanedione(trigger)

Mix 22 hours(G2.3) Shaking 1 hr

2.5mg2,6-di-tert-butyl-4-methylphenol+10% Diox.

Anti-poly60C (G1.3)

9.3ulHydrogen Peroxide12.5mgAntarctica lipase (free)+ 40% Diox.

Enzyme(G1.2)

2 ul waterDissolve(G2.2)

1.5mlx2anhydrous Dioxane

1.6mg/

0.05ml

HRP

0.11 mlTetrahydrofuran (solvent)(THF) N2 flushed

0.182 mL, 1.278 mM

2,2,2-trifluoroethyl methacrylate

~0.02 g 0.082 mM

L-Ascrbyl methylmethacrylate

Mix1(G2.1) w/G2.2

150mg, 0.852 mM

L-ascorbic acid(AA)+50% Diox.

FunctionalSubstrate(G1.1)

Quantityused

MaterialStepQuantity used

MaterialStep

Stage II HRP Polymerization L-Ascrbyl Methylmethacrylate

Reaction vessel 260 min.<reaction time<90 min. 20 min.<shaking time<30 min.Flow rate<0.01 ml/min.

2Stage I Enzymatic TransesterificationSynthesis L-Ascrbyl Methyl methacrylate

Reaction vessel 150C<reaction temp <60C, 45 min.<reaction time<60 min.Flow rate<0.01 ml/min.

1

1st Vessel>50C

2nd Vessel

Hydrogenperoxide

2,6-di-tert-butyl-4-methylphenol, Dioxane.

A

BC

D

E

Initiator

HRP,THF

Ascorbicacid, Dioxane

TFM, Diox.Lipase,

AA_PMMA,/PMMA/

Function driven


ASIM manufacturingASIM manufacturing

• DRIE Si wafer• PDMS Casting• Thermal Plasma Bonding to

glass slide• Embed fluid connectors

PDMS on Glass slide

PDMS on SI


External HardwareExternal Hardware

Micrometer

Sample loading

Pneumatic controlling

Syringes


Chemical AnalysisChemical Analysis

Repeat unit

Repeat unit

Molecular weight/ charge

Signal strength

Macro

Micro

• Macro vs. Micro comparison with MALDI-TOF• Need purification but polymer exists


TeamTeam

• David Kaplan - expertise in enzymatic reactions

• Peter Wong - expertise in microfluidics

• Jin Zou - PhD graduate in Mechanical Engineering

• Martin Son - Tufts Technology Transfer Office

• Tufts Capabilities:– Enzymatic synthesis research, development, and

production

– ASIM - Microfluidic design, analysis, and fabrication

– Polymer discovery program – design of experiments and testing


Next StepsNext Steps

• Identify 2 to 3 market products to tackle– 2 months

• Initial description of enzymatic synthesis process– 2 months

• Convert preliminary patent application to full application with these examples of synthesis– 1 month

• Develop next generation of ASIM devices for those specific market products– 6 months

• Develop new polymer products– 6 months

• Partner with companies to develop new polymers for their markets


SummarySummary

• Food/Medicine/Biochem Markets need advantages of new polymers

• Microfluidic Enzymatic Synthesis– Make custom polymers– Faster, cheaper discovery– Scalable to mass production

• Need partners and funding to – do market analysis,– help secure IP, – develop small number of prototypes, and – expand to market

• Contact [email protected]

Office for Technology Licensing and Industry Collaboration ● Tufts School of Engineering Polymer...

Documents

Transcript of Office for Technology Licensing and Industry Collaboration ● Tufts School of Engineering Polymer...