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NanobioprocessingNanobioprocessing Larry P. Walker, Ph.D. Professor, Department of Biological and Environmental Engineering Cornell University

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Agricultural and Environmental Bioprocess Engineering 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

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

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Nanobiotechnology is beginning to generate substantial new insights into how biological systems function, and likewise, nanobiotechnology will lead to the design of entirely new classes of micro- and nanofabricated devices and systems. Nanobiotechnology Center (NBTC) Cornell University

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Nanobiotechnology Center (NBTC) Cornell University Cornell University Clark Atlanta University Howard University Princeton University Oregon Health Sciences University Wadsworth Center New York Department of Public Health

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

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

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

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Membranes and Packings Semipermeable Membranes and Equilibrium Dialysis Michael Spencer, Lori Lepak, Jim Turner, Michele Caggana Fabrication of Integratable Semipermeable Membranes Jim Turner, Peter Russo, David Martin, David Lawrence, Michele Caggana, Bill Shain, Michael Isaacson, and Michael Spencer High Resolution Protein Purification using Block Copolymer Derived Mesoporous Silica-type Materials Uli Wiesner, Phong Du, Caroline Corner, & Larry Walker Porous Block Copolymers For Microfluidic Separation Chris Ober &Mingqi Li

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Micro and Nanofluidic devices Micro and Nanofluidics for Molecular Separation and Detection Harold Craighead, John Henion, David Czaplewski, Yanou Yang & Jun Kameoka Multidimensional Microscale Separations of Cerebrospinal Fluid Proteins Kelvin Lee, Michael Shuler, Jim Engstrom, Bob Austin, Chen Li & Jinpian Diao Nanostructures for Enzyme Transport and Accessibility Assessment Larry Walker, Harold Craighead, Jennifer Guisado & Tina Jeoh

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Biomolecular Devices and Analysis Some Common Activities: Lithography Chemical synthesis Spinning of molecules Assessing morphology Assessing monodispersity Assessing transport mechanisms Measurement & Analysis

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Biomolecular Devices and Analysis Biocompatibility, Working with small volumes: pico- to micro-liters, Sieving and sorting at the scale of 5, 10, 15 and 50 nm, Monodispersity tight control over pore size distribution and shape

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Example: Membrane Technology Si Si3N4 Spun Organic membrane Spin Coating of Cellulose And Collagen onto silicon surfaces to create porous membranes

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Fabrication of Integratable Semipermeable Membranes M. Spencer 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

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Block Co-polymers Uli Wiesner & Chris Ober 12 hrs 400V/cm (b) (c) ChemicalSynthesis PhotoLithography excellent biocompatibility of silica easily accessible pore sizes from 8-50 nm narrow pore size distributions

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Nanostructures for Enzyme Transport and Accessibility 1.Studying cellulase binding and catalysis as a function of molecular diameter. 2.Study synergism in cellulase/ cellulose reaction systems.

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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 m Multidimensional Microscale Separations Proteins K. Lee

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Micro and Nanofluidics for Molecular Separation and Detection H. Craighead Bottom Plastic Chip Embossed Top Plastic Chip Reservoir hole Embossed Fluidic Channel Emitter filmBonding Electrospray Ionization

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(a) (b) Triangle Nozzle Taylor cone Liquid jet Plume 50 m Berberine:FW=336 (a) (b) Application to Mass Spectrometry Analysis

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NBTC At the Interface between Biology and Engineering Nanobiotechnology Advanced Bioreactors Advanced Energy Systems Controlled Environment Agriculture Systems Genomics Proteomics Metabolic Engineering Protein Engineering

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We are having fun!