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Nanoporous Membrane Technologies for Pathogen Collection, Separation, and

DetectionSang Won Lee, Hao Shang, and Gil U Lee*

School of Chemical Engineering, Forney Hall, Purdue University, West Lafayette, IN 47906

and

Matthew T. Griffin and Jack FultonNVEO & Chem/Bio Sensors Group

NSWC CraneCrane, IN 47522

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4. TITLE AND SUBTITLE Nanoporous Membrane Technologies for Pathogen Collection,Separation, and Detection

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Standard Form 298 (Rev. 8-98) Prescribed by ANSI Std Z39-18

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Approach

Develop membranes and methodology for continuous collection of airborne particles.

Schematic of a Point Detector that Utilizes Membrane

Develop a membranes and and methodology for continuous separation via ultrafiltration.

Develop a receptor functionalized membrane to improve mass transport and kinetic conditions.

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Nanoporous Membranes! Nanoporous alumina membranes were chosen as a substrate because of their desirable physical properties and high density of uniform pores of 10-200 nm size.

! The membrane surfaces will be modified with hydrophobic and hydrophilic coatings to facilitate pathogen collection and separation.

! The membrane surfaces will be modified with proteins and nucleic acids to enable pathogen identification. Side view: 100 nm

pore, 100 micron thick alumina membrane

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Membrane Chemistries

PEI-PEG (2000)O

H

OH

OH

OH

OH

5 % PEI, pH=8.2

NH

2

NH

2

NH

2

NH

2

NH

2

M-PEG, pH=8.2

OC

H3

OC

H3

OC

H3

OC

H3

OC

H3

OHOHOH

Si

CH3O

CH3O

CH3O OCH3+

OOO

Si OCH3

Reflux in toluene w/ 5%Et3N for 4 hrs under N2

Silane-PEG (5000)

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Characterization of Membrane Chemistries

15.3

38.6

11.0

35.8

58.4

% O 1s (530.3 eV)

21.05.650.29.05.0PEI-PEG

35.81.224.5OTMS coating

21.83.446.42.111.0Silane-PEG

12.233.219.2PEI

11.130.3Unmodified

membrane

% O 1s (532.0 eV)

% N 1s (399.1 eV)

% C 1s (286.5 eV)

% C 1s (285.6 eV)

% Si 2p (103.5 eV)

% Al 2p (74.4 eV)

Chemistry

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Protein Fouling

1000

800

600

400

200

0

Flu

ore

scen

eIn

ten

sity

(a.

u)

2.01.51.00.50.0

BSA Concentration (mg/ml)

Silane-PEG coated Membrane

Unmodified Membrane

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Collector

Air in

To pump Membrane holder Flow meter

Relative humidity meter

HumidityControl

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Gas and Liquid Permeabilities

Nitrogen permeability (m/PaS)

Water permeability (m/PaS)

6.84E-06

-

2.03E-06

Silane-PEG

6.88E-6

6.08E-06

1.90E-06

--6.50E-06200 nm

3.14E-064.63E-064.62E-06100 nm

7.08E-082.0E-061.98E-0620 nm

PEGPEIBare membraneHolder 2 Holder 1

2.73E-084.06E-103.04E-092.18E-082.86E-08200 nm

-Foulingn/a2.14E-081.91E-08100 nm

6.59x10-09Fouling1.64E-092.67E-097.00E-0920 nm

Silane-PEGPEGPEIBare membraneHolder 2 Holder 1

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Membrane Permeabilities in the Presence of Water

0

100

200

300

400

500

600

700

0 5 10 15 20 25 30

Pressure (PSIG)

Flo

w R

ate

(SC

CM

)

Dry Air

0.028 H2Omolefraction

0.018 H2Omolefraction

0.010 H2Omolefraction

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Operation Characteristics

0After filtration

2x106Before

BG spores

(spores/ml)

Collection Efficiency

96.7Sonication in PBS

26.9H2O

76.4SDS 10 %

75.4PBST 0.5 %

Removal Efficiency

(%)

Method of Extraction

Extraction Efficiency

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3 Day Laboratory Trial

10x10-6

8

6

4

2

0

Per

mea

bilit

y (m

/Pa.

s)

706050403020100Time (hr.)

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Collector Prototype

Air In

PressureSensor

Membrane

Membraneholder

Quick-disconnect

Inlet Cell

Quick-disconnect

Ultrasonic Source

Quick-disconnect In

Out

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Performance of the Prototype

0

10

20

30

40

50

60

70

250000.00 350000.00 450000.00 550000.00 650000.00 750000.00 850000.00

Pressure (Pa)

Flo

w R

ate

(lit

res/

min

)

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Separator - Solute Permeabilities

20nm membrane

3.5

3.0

2.5

2.0

1.5

1.0

0.5

Ab

s.

12001000800600400200

Time (min)

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Protein Permeability

7x10 -708.19x10 -76.37x10-7

Silane-PEGPEI-PEGUnmodified Membrane

Theoretical Permeability of Ovalbumin

(cm2/s)

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Collector & Separator Fluidics

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Detection

Adsorption Scattering

Patent sensitive

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Self-assembled Monolayers

Atomic force microscope image of a 12 nm Au array. Scan size 500 x 500 nm. Z scale 50 nm

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Ovalbumin Binding

0.34

0.32

0.30

0.28

0.26

0.24

0.22

0.20

Abs

orba

nce

800700600500400Wavelength (nm)

A)

B)

Patent sensitive

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Nanoporous Membrane Separation Methodologies

Point of Contact: Gil Lee

School of Chemical EngineeringPurdue University

West Lafayette, IN 47907-1283765-494-0492

gl@atom.ecn.purdue.edu

Objective: To fabricate and characterize the performance nanoporous membranes for the collection, separation, and detection of airborne pathogens.

Payoff: A highly sensitive point detector that consumes minimal reagents. Design criteria include < 0.1 ACPLA sensitivity for toxins, viruses and bacteria; < 10 min response time; < 0.5 ml/min total reagent consumption.

Top view:100 nm pore alumina membrane

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Acknowledgements

Helpful discussionGavin Reid and Scott McLuckey, Dept. Chemistry, Purdue UniversityNorman Hovijitra, Eric Wallis, and Richardo Chong, School of Chemical EngineeringJoe Brumfield, ONRRichard Haash, UIUCJerry Bottiger, SBBCOM

FundingInstitute for the Detection of Hazardous Materials Purdue University, ONR, and the Shreve Trust.