Nanobioprocessing

NanobioprocessingNanobioprocessing

Larry P. Walker, Ph.D.Larry P. Walker, Ph.D.Professor, Department of Biological and Professor, Department of Biological and Environmental EngineeringEnvironmental EngineeringCornell UniversityCornell University

Agricultural and Environmental Bioprocess Agricultural and Environmental Bioprocess Engineering ResearchEngineering Research

Molecular mechanisms of polysaccharide degrading enzymes,Solid state microbial processes and molecular ecology,Mechanisms and kinetics of metal uptake and sequestration in plants,Molecular filtration and analysis of biomolecules

Nanobiotechnology Center (NBTC)Nanobiotechnology Center (NBTC)Cornell UniversityCornell University

““The Nanobiotechnology Center (NBTC) was The Nanobiotechnology Center (NBTC) was established in January 2000 as a Science & Technology established in January 2000 as a Science & Technology Center, with core funding from the National Science Center, with core funding from the National Science Foundation. Nanobiotechnology is an emerging area of Foundation. Nanobiotechnology is an emerging area of scientific and technological opportunity that integrates scientific and technological opportunity that integrates nano/microfabrication and biosystems to the benefit of nano/microfabrication and biosystems to the benefit of both.” both.”

www.nbtc.cornell.eduwww.nbtc.cornell.edu

““Nanobiotechnology is beginning to generate Nanobiotechnology is beginning to generate substantial new insights into how biological systems substantial new insights into how biological systems function, and likewise, nanobiotechnology will lead function, and likewise, nanobiotechnology will lead to the design of entirely new classes of micro- and to the design of entirely new classes of micro- and nanofabricated devices and systems.”nanofabricated devices and systems.”




Cornell UniversityCornell UniversityClark Atlanta UniversityClark Atlanta UniversityHoward UniversityHoward UniversityPrinceton UniversityPrinceton UniversityOregon Health Sciences UniversityOregon Health Sciences UniversityWadsworth Center – New York Department of Wadsworth Center – New York Department of Public HealthPublic Health



Cellular DevicesCellular Devices (David Lawrence, Wadsworth Center)– (David Lawrence, Wadsworth Center)– studies of engineered devices for separating and studies of engineered devices for separating and analyzing individual cells.analyzing individual cells.Nanoscale Cell BiologyNanoscale Cell Biology (Manfred Lindau, Cornell (Manfred Lindau, Cornell University)University) - micro- and nanofabricated tools to - micro- and nanofabricated tools to understand and utilize cellular responses on the understand and utilize cellular responses on the nanoscale.nanoscale.Biomolecular DynamicsBiomolecular Dynamics (Michael Koonce, Wadsworth (Michael Koonce, Wadsworth Center) –Center) – exploration and utilization of molecular motion exploration and utilization of molecular motion and mechanical properties and mechanical properties



Cell-Surface InteractionsCell-Surface Interactions (Harvey Hoch, Cornell (Harvey Hoch, Cornell University) – University) – Investigating the response and control of Investigating the response and control of cellular responses on structured surfacescellular responses on structured surfacesBiomolecular Devices and Analysis Biomolecular Devices and Analysis (Larry Walker, Cornell (Larry Walker, Cornell University)University) - investigation new approaches to high speed - investigation new approaches to high speed and highly sensitivity analysis of biomolecules.and highly sensitivity analysis of biomolecules.Nanoscale Biomaterials Nanoscale Biomaterials (Dotse Sogah, Cornell University)(Dotse Sogah, Cornell University) – study of materials engineered at the nanoscale for new – study of materials engineered at the nanoscale for new properties that can be exploited in nanobiotechnolgy.properties that can be exploited in nanobiotechnolgy.

Major research and Major research and development activities in development activities in the life sciences has the life sciences has generated the need for generated the need for materials, methods, and materials, methods, and devices for sorting, devices for sorting, separating, and analyzing separating, and analyzing proteins, DNA and other proteins, DNA and other biomolecules. biomolecules.

Biomolecular Devices and AnalysisBiomolecular Devices and Analysis

Membranes and PackingsMembranes and PackingsSemipermeable Membranes and Equilibrium DialysisSemipermeable Membranes and Equilibrium Dialysis

Michael Spencer, Michael Spencer, Lori LepakLori Lepak, Jim Turner, Michele , Jim Turner, Michele CagganaCaggana

Fabrication of Integratable Semipermeable MembranesFabrication of Integratable Semipermeable MembranesJim Turner, Jim Turner, Peter Russo, Peter Russo, David Martin, David Lawrence, David Martin, David Lawrence,

Michele Caggana, Bill Shain, Michael Isaacson, and Michele Caggana, Bill Shain, Michael Isaacson, and Michael SpencerMichael Spencer

High Resolution Protein Purification using Block High Resolution Protein Purification using Block Copolymer Derived Mesoporous Silica-type MaterialsCopolymer Derived Mesoporous Silica-type Materials

Uli Wiesner, Phong Du, Caroline Corner, &Uli Wiesner, Phong Du, Caroline Corner, & Larry WalkerLarry Walker

Porous Block Copolymers For Microfluidic SeparationPorous Block Copolymers For Microfluidic SeparationChris Ober &Chris Ober &Mingqi LiMingqi Li

Micro and Nanofluidic devicesMicro and Nanofluidic devices Micro and Nanofluidics for Molecular Separation and Micro and Nanofluidics for Molecular Separation and

DetectionDetectionHarold Craighead, John Henion, David Czaplewski, Harold Craighead, John Henion, David Czaplewski,

Yanou Yang & Jun KameokaYanou Yang & Jun Kameoka

Multidimensional Microscale Separations of Multidimensional Microscale Separations of Cerebrospinal Fluid ProteinsCerebrospinal Fluid Proteins

Kelvin Lee, Michael Shuler, Jim Engstrom, Bob Austin, Kelvin Lee, Michael Shuler, Jim Engstrom, Bob Austin, Chen Li & Jinpian DiaoChen Li & Jinpian Diao

Nanostructures for Enzyme Transport and Accessibility Nanostructures for Enzyme Transport and Accessibility AssessmentAssessment

Larry Walker, Harold Craighead, Jennifer GuisadoLarry Walker, Harold Craighead, Jennifer Guisado& Tina Jeoh& Tina Jeoh

Biomolecular Devices and AnalysisBiomolecular Devices and Analysis

Some Common Activities:Some Common Activities: LithographyLithography Chemical synthesisChemical synthesis Spinning of moleculesSpinning of molecules Assessing morphologyAssessing morphology Assessing monodispersityAssessing monodispersity Assessing transport Assessing transport

mechanismsmechanisms Measurement & Measurement & AnalysisAnalysis

Biomolecular Devices and AnalysisBiomolecular Devices and AnalysisBiomolecular Devices and AnalysisBiomolecular Devices and Analysis

Biocompatibility,Biocompatibility,

Working with small volumes: pico- to Working with small volumes: pico- to micro-liters,micro-liters,

Sieving and sorting at the scale of 5, 10, Sieving and sorting at the scale of 5, 10, 15 and 50 nm,15 and 50 nm,

Monodispersity – tight control over pore Monodispersity – tight control over pore size distribution and shapesize distribution and shape

Example: Membrane TechnologyExample: Membrane TechnologyExample: Membrane TechnologyExample: Membrane Technology

Si

Si3N4

SpunOrganicmembraneSpin Coating of CelluloseSpin Coating of Cellulose

And Collagen onto And Collagen onto silicon surfaces to create silicon surfaces to create porous membranes porous membranes

Fabrication of Integratable Semipermeable Fabrication of Integratable Semipermeable Membranes – M. Spencer Membranes – M. Spencer

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1 layer collagen:Methyl orange (MW 327) diffuses through in 10 min

4 layers collagen:No

methyl orange diffuses in 10 hours (up to 48 hours!)

400 nm thick

Pores 0-100nm

Collagen

Dyes of 270 –

390 Da

Block Co-polymersBlock Co-polymersUli Wiesner & Chris OberUli Wiesner & Chris Ober

Block Co-polymersBlock Co-polymersUli Wiesner & Chris OberUli Wiesner & Chris Ober

12 hrs12 hrs

400V/cm400V/cm

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ChemicalChemicalSynthesisSynthesis

PhotoPhotoLithographyLithography

excellent biocompatibility excellent biocompatibility of silicaof silica

easily accessible pore easily accessible pore sizes from 8-50 nmsizes from 8-50 nm

narrow pore size narrow pore size distributionsdistributions

Nanostructures for Enzyme Transport and AccessibilityNanostructures for Enzyme Transport and Accessibility

1.1. Studying cellulase binding Studying cellulase binding and catalysis as a function and catalysis as a function of molecular diameter.of molecular diameter.

2.2. Study synergism in Study synergism in cellulase/ cellulose reaction cellulase/ cellulose reaction systems.systems.

Channel design & fabrication

Make inlets and outlets

Glass-Glass bonding

Attach peripheral components

Flow chart of the device fabrication process

New 2 stage bonding approach developed to permit alignment and

bonding.

GMA-co-TRIM polymer

Double-T Structure

Channel with affinity separation bed

5 μ

m

Multidimensional Microscale Separations ProteinsMultidimensional Microscale Separations Proteins K. LeeK. Lee

Micro and Nanofluidics for Molecular Separation Micro and Nanofluidics for Molecular Separation and Detection – H. Craighead and Detection – H. Craighead

Bottom Plastic Chip

Embossed Top Plastic Chip

Reservoirhole

Embossed Fluidic Channel

Emitter film Bonding

Electrospray IonizationElectrospray Ionization

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Taylor cone

Liquid jet

Plume

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CH3O

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Application to Mass Application to Mass Spectrometry AnalysisSpectrometry Analysis

NBTC At the Interface between Biology and NBTC At the Interface between Biology and EngineeringEngineering

• Nanobiotechnology• Advanced Bioreactors• Advanced Energy Systems• Controlled Environment

Agriculture Systems

•Genomics•Proteomics•Metabolic Engineering•Protein Engineering

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Nanobioprocessing