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

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

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Polymer Discovery Via Polymer Discovery Via Microfluidic Enzymatic SynthesisMicrofluidic Enzymatic Synthesis

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

October 3, 2006

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OverviewOverview

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

Biochemical Science

Enzyme Polymer science 1970-

Synthesis 1990-

Micro-fabrication

Microsystems Technology

Engineering

Mechanical Electronics

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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

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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

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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?

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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.

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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

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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

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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

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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

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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

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ASIM manufacturingASIM manufacturing

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

glass slide• Embed fluid connectors

PDMS on Glass slide

PDMS on SI

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External HardwareExternal Hardware

Micrometer

Sample loading

Pneumatic controlling

Syringes

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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

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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

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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

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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 Martin.Son@tufts.edu