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Nanofluidic Microsystems for Advanced Biosample Preparation

Microfluidics Tech Fair 2006. Nanofluidic Microsystems for Advanced Biosample Preparation . Ying-Chih Wang (ycwang@mit.edu) 1 , Jianping Fu, Yong-Ak Song and Jongyoon Han 2,3

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Nanofluidic Microsystems for Advanced Biosample Preparation

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  1. Microfluidics Tech Fair 2006 Nanofluidic Microsystems for Advanced Biosample Preparation Ying-Chih Wang (ycwang@mit.edu)1, Jianping Fu, Yong-Ak Song and Jongyoon Han 2,3 1Department of Mechanical Engineering, 2Biological Engineering Division, 3Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, MA 02139 October 3rd, 2006

  2. The need and the market for sample preparation • Market of Proteomics, $1.3 billion and growing (13% annually) • Greatest challenge in proteomics • Sample complexity (>20,000 different proteins)  Purification required • 2D gel electrophoresis, $800 M in 2004 ($1.8B 2011) • Time and labor consuming • Poor recovery for low abundance sample after multiple steps • Consumers: Biologist, Pharmaceutical R&D, clinical diagnostics IN Detection Fraction Separation (Mass Spectrometry) (2D gel analysis)

  3. + + - - - + + - Microchip Our Approach (gel free) Complex peptide/protein mixture Sample preparation in microfluidic chip Size-based separation Charge-based separation Preconcentration Sensing/Detection Droplet/Electrospray

  4. Our Core (Patented) Techniques • Nanofluidic molecular sieving Continuous biomolecule size separation • Microfluidic charge-based sorting Continuous biomolecule charge separation • Electrokinetic nanofluidic preconcentrator Rapid molecular trapping Enable rapid and economical low-abundant sample identification

  5. Fabrication Method of Nanofluidic Devices Fabrication DO NOT need nanolithography Thin channel instead of Narrow channel Uniform, flat nanofluidic channel confirmed down to 20nm Making nanofluidic (20 nm) device using standard microfabrication techniques! Pan Mao and Jongyoon Han, 2005, EECS / BE / MIT

  6. Core Technology I:Nanofluidic filter array for size-based separation

  7. Molecular Separation in 2-D Nanofilter Arrays • Physically hinders protein migration • Continuous two- dimensional separation

  8. Continuous flow separation video (All 20 Speed) Ogston sieving,DNA (50 bp – 766 bp, 5 bands) Ogston sieving, protein complex (11 kDa vs. 116 kDa) Entropic trapping, DNA (2 kbp – 23 kbp, 6 bands) 1960 µm 4080 µm 1960 µm Small Large Size: Large Small Large Small •  DNA - Hind III digest. • Ex=380 V/cm, Ey=400 V/cm • SDS-Protein complex. • Ex=150 V/cm, Ey=200 V/cm. • PCR marker. • Ex=70 V/cm, Ey=100 V/cm • A general but unique size-based separation tool. Even larger DNA (~Mbp) possible with this method. J. Fu, A. Stevens, S. R. Tannenbaum & J. Han. subminted

  9. Core Technology II:Charge separation driven by diffusion potential (no external power) c=200mM c=1mM • Different diffusivities of the buffer ions • generate a diffusion potential across the • liquid junction • Potential gradient (electric field) utilized • for binary sorting

  10. Ampholyte-free pI-based separation Continuous-flow operation Song, Y.-A., Hsu, S., Stevens, A.and Han, J. Anal. Chem. (2006).

  11. 20 mm Core Technology III:Preconcentration by Ion Selective Nanofluidic Channels Electrical double layer overlapping: Device layout: Wang et al, Anal. Chem., 77, 4293 micro channel: cross section 1x10mm ~50mmx50mm length 1 -2 cm nano channel: cross section 40nm x20mm ~40mmx200mm length 100 -200mm Allen, Bard “Electrochemical Methods” Fabrication: Mao et al.,  Lab Chip, 2005, (8),837-844

  12. t = 0 t = 40 min Preconcentration Mechanism ET ( )

  13. Million-fold Protein Concentration Enhancement 107 105 Wang, Y.-C., Stevens, A. L.and Han, J. Anal. Chem. 77, 4293-4299 (2005). Regular, stable pore size contributes its long term stability

  14. size-based Sorter pI-based Sorter Vision: The Integrated sample preparation device Protein Concentrator Silicon-based technologies make integration easier Charge Separation Size separation Pre-concentration

  15. Developing Timeline Our advantages: • Rapid biomolecule separation (<30 mins) • Minimum sample consumption (<1 ml) • Automated solution for biomarker discovery/ tracking • Better recovery compares to 2D gel (no post processing) • Direct coupling to mass spectroscopy or immunoassay Principal Investigator: Jongyoon (Jay) Han, jyhan@mit.edu

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