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Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005 For Online Review Only Kenneth Carter UMASS 2005 Biopatterned Surfaces Carter Research Group University of Massachusetts
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Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

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Page 1: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

BiopatternedSurfaces

Carter Research GroupUniversity of Massachusetts

Page 2: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

NanoMolding – embed functionality into feature- use LFRP to incorporate functionality

NanoMolding – embed functionality into feature- use LFRP to incorporate functionality

Protein chipsProtein chips

Biopatterned Surfaces

Page 3: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

B B B B B

B B B B BB

B

Biotin patterned surfaceStreptavidin

Incubation in aq. soln. of avidin

Wash away excess avidin and incubate with a biotin

conjugate of the biomolecule

BBiotinylated Biomolecule( e.g., peptides, DNA, or

proteins )

B B B B BB

B B B BB BSurface patterned withbiomolecules

=

=

• Qi, K.; Wooley, K. L.; Jhaveri, S. B.; Sogah, D. Y.; Beinhoff, M.; Malkoch, M.; Carter, K. R.; Hawker, C. J. PMSE Prep., 2004, 91133-134.

• Hawker, C. J.; Wooley, K. L.; Carter, K. R.; Sogah, D. Y.; Qi, K.; Jhaveri, S. B.; Beinhoff, M.; Malkoch, M. PMSE Prep., 2004, 90, 279.

Biopatterned Surfaces

Page 4: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005O N

X

O N

.+

O N

X

.

.+ O N.

120o C

120o C

Exchange between nitroxides

Alkoxyamine within crosslinked polymer

Alkoxyamine within crosslinked polymer

( Large excess )

( very small %) O

Functional groupexchanged onto

the growing brush ends

Functional groupexchanged onto

the growing brush ends

Nitroxide Exchange:- Based on equilibrium between dormant alkoxyamine and active nitroxide

Added ‘free’ functional alkoxyamine in solution

Added ‘free’ functional alkoxyamine in solution

PolyO N

X

Functionality via Nitroxide Exchange

Page 5: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

O N

X

Polymerize brushes in the presence of functional

alkoxyamine

Polymerize brushes in the presence of functional

alkoxyamine

X= Desired Functional Groupto be introduced on the surface

X XXXX X

XX

XX

XXX XXXX X

XX

XX

XX

X XXXX X

XX

XX

XX

Exchanged Functional groupExchanged Functional group Polymer brushes – control solvationPolymer brushes – control solvation

Embedded Alkoxyamine InitiatorsEmbedded Alkoxyamine Initiators

Functionality via Nitroxide Exchange

Page 6: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

Example: Biotinylated Surfaces

Example 1: Biotin - A Biological Ligand

• Biotin functionality is covalently attached to the “nitroxide”part of the alkoxyamine

O N

OH

O N

Br

NaN3

DMF

O N

N3

O N

NH2

LiAlH4

THF

Biotin-NHS esterO N

NH

OS

NH NH

O

PPh3

DMF, Et3N

CBr4

HH

• Qi, K.; Wooley, K. L.; Jhaveri, S. B.; Sogah, D. Y.; Beinhoff, M.; Malkoch, M.; Carter, K. R.; Hawker, C. J. PMSE Prep., 2004, 91 133-134.

• Hawker, C. J.; Wooley, K. L.; Carter, K. R.; Sogah, D. Y.; Qi, K.; Jhaveri, S. B.; Beinhoff, M.; Malkoch, M. PMSE Prep., 2004, 90, 279.

Page 7: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

B BBBBB

BB

BB

BBB BBBBB

BB

BB

BB B BBBBB

BB

BB

BB

125o C

DMF

Silicon / Quartz substrate

O NO NMe2 .O N

NH

OS

NH NH

O

HH

1.168.75 KSilicon1

PDIMnSub.Run#

Alexa Flor labelledstreptavidin

10 mM HEPES buffer

Scale bar = 100um

PDMA no Biotin

Biotinylated PDMA

Bioresponsive Surfaces

• Qi, K.; Wooley, K. L.; Jhaveri, S. B.; Sogah, D. Y.; Beinhoff, M.; Malkoch, M.; Carter, K. R.; Hawker, C. J. PMSE Prep., 2004, 91 133-134.

• Hawker, C. J.; Wooley, K. L.; Carter, K. R.; Sogah, D. Y.; Qi, K.; Jhaveri, S. B.; Beinhoff, M.; Malkoch, M. PMSE Prep., 2004, 90, 279.

Page 8: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

Scale bar = 100um

40 um

Selective attachment of labeled streptavadin!

Individual 100 nm lines could not be resolved due to limitations of confocal

microsxopy

Selective attachment of labeled streptavadin!

Individual 100 nm lines could not be resolved due to limitations of confocal

microsxopy

100 um

Fluorescence Images

Qi, K.; Wooley, K. L.; Jhaveri, S. B.; Sogah, D. Y.; Beinhoff, M.; Malkoch, M.; Carter, K. R.; Hawker, C. J., Manuscript in Preparation, 2005

Page 9: Carter Research Group University of Massachusettskcarter/research/biopatterned.pdf · BBBBBB B B B B B B BBBBBB B B B B B B BBBBBB B B B B B B 125 o C DM F Silicon / Quartz substrate

Page from Carter Group Website - UMass For online viewing only – do not print, copy or redistribute Copyright Carter 2005

For Online Review Only

Kenneth CarterUMASS 2005

IgE / Dinitrophenol Interactions

(1) First IgETreatment

(2) DNP-BSA HaptenDetachment Treatments

10 um10 um

40 um

10 um

(3) Second IgETreatment

10 um

Qi, K.; Wooley, K. L.; Jhaveri, S. B.; Sogah, D. Y.; Beinhoff, M.; Malkoch, M.; Carter, K. R.; Hawker, C. J., Manuscript in Preparation, 2005