Polymer Discovery Via Microfluidic Enzymatic Synthesis

1
Polymer Discovery Via Microfluidic Enzymatic
Synthesis

• Prof. Peter Y. Wong
• Prof. David Kaplan
• Tufts University - Medford, MA
• October 3, 2006

2
Overview

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

3
Many Markets

• Any market that benefits from new biochemicals
• Improved Foods
• Additives, Modification, Nutrition
• Green Chemistry
• Agricultural, Packaging, Analysis
• New Medicines
• Topical, Digested, Structural

4
Markets Needs vs.Wants

• New products
• Better
• Faster
• Cheaper
• Differentiated
• Macro and Micromolecules needed
• New material
• New processes
• Focus on new polymers and processes

5
Problems 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?

6
Our 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.

7
Microfluidic 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.

8
PMMA 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

9
ASIM

• Goals
• Two enzymatic cascade reactions to produce PMMA
• low cost devices made of poly(dimethylsiloxane)
  (PDMS)
• efficient method to optimize process with
  external controls

10
Translation from Abstract to Hardware
monomer
ascorbic acid
AA-Monomer
AA- Ascorbic Acid MMA- Methyl Methacrylate PMMA-
Poly (Methyl Methacrylate) P-AA-MMA Ploy
L-Ascorbic Methyl Methacrylate HRP Horse Radish
Peroxidase
lipase
hydrogen
HRP
peroxide
ascorbic acid
Antioxidant polymer
monomer
Input

Input

Input
hydrogen
Input
ascorbic acid
ascorbic acid
HRP
peroxide


1
2
Input

Input

Check Valve
Output
Output
Reaction Vessel 1
Reaction Vessel 2
Reaction Vessel 1
monomer
polymer
in solvent
in solvent
AA-Monomer
P-AA-MMA
AA-Monomer
React with lipase
React with
HRP
Output

unreacted
Output

unreacted
Hydrogen peroxide
Output

unreacted
Output

unreacted
lipase
hydrogen peroxide
hydrogen peroxide
ascorbic acid
ascorbic acid
11
Improved Version
Stage II HRP Polymerization L-Ascrbyl
Methylmethacrylate Reaction vessel 2 60
min.ltreaction timelt90 min. 20 min.ltshaking
timelt30 min. Flow ratelt0.01 ml/min.
2
Stage I Enzymatic Transesterification Synthesis
L-Ascrbyl Methyl methacrylate Reaction vessel
1 50?Cltreaction temp lt60?C, 45 min.ltreaction
timelt60 min. Flow ratelt0.01 ml/min.
1
Function driven
Quantity used
Material
Step
Quantity used
Material
Step
2,6-di-tert-butyl-4-methylphenol, Dioxane.
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.
Functional Substrate (G1.1)

Ascorbic acid, Dioxane
1st Vessel gt50?C
0.11 ml
Tetrahydrofuran (solvent) (THF) N2 flushed
0.182 mL, 1.278 mM
2,2,2-trifluoroethyl methacrylate
A
Initiator
B
1.6mg/ 0.05ml
HRP
2nd Vessel
C
TFM, Diox. Lipase,
2 ul
water
Dissolve (G2.2)
1.5mlx2
anhydrous Dioxane
D
HRP,THF
9.3ul
Hydrogen Peroxide
12.5mg
Antarctica lipase (free) 40 Diox.
Enzyme (G1.2)
Hydrogen peroxide
E
AA_PMMA, /PMMA/
1.77ul
2,4-pentanedione (trigger)
Mix 2 2 hours (G2.3) Shaking 1 hr
2.5mg
2,6-di-tert-butyl-4-methylphenol 10 Diox.
Anti-poly 60?C (G1.3)
Vessel 2 Poly L-Ascorbyl Methyl
methacrylate (P-AA-MMA) (G.2)
Vessel 1 L-Ascrbyl Methyl methacrylate (AA-MMA)
(G 1)
12
ASIM manufacturing

• DRIE Si wafer
• PDMS Casting
• Thermal Plasma Bonding to glass slide
• Embed fluid connectors

PDMS on SI
PDMS on Glass slide
13
External Hardware
14
Chemical Analysis

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

15
Team

• 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

16
Next 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

17
Summary

• 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_at_tufts.edu