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Microfabricated Nanofluidic Sieving Structures for Rapid Biomolecule Separation
Jianping Fu, Hansen Bow, Pan
Mao, Reto B. Schoch
is a fundamental analytical and preparative technique
in biology, medicine, chemistry, and industry. Fractionation
of biological molecules, such as nucleic acids and proteins,
plays a central role in genomic analysis. In the new challenge
of systems biology, as well as in the application of biomarker
detection and biosensing, this task becomes even more important
because solving the puzzle of interactions between proteins
is far more complicated than deciphering genomes, due to the
lack of protein's equivalent of amplification, fractionation
and sequencing techniques.
project seeks to use microfabricated regular nanofluidic filters
(nanofilters) as controllable sieving medium for size- or charge-based fractionation
of various biologically relevant macromolecules, such as ds
DNA, proteins, and polysaccharides. Using standard microfabrication
techniques, we have precisely fabricated nanofilters with
gap thickness down to the vicinity of 10 nm. In such molecular-scale
confining structures, molecular transport properties are largely
affected by the steric constraints of nanofluidic structures.
this project, we construct different nanofilter based
separation devices and explore the steric and electrostatic partitioning
of biomolecules with the nanofilter for size- or charge-fractionation
of various biologically relevant macromolecules. In the proposed
separation systems, unlike other conventional random nanoporous
sieving materials, the nanofilters can be made uniform and
controllable, also chemical groups on the wall can be tailored.
In addition to the application of biomolecule separation,
the nanofilter based artificial sieving structures provide
an ideal platform for the theoretical study of molecular dynamics
and stochastic motion in confining spaces because of their
precisely characterized environments.
One-Dimensional Periodic Nanofilter Array for Biomolecule Separation. (Jianping Fu, Pan Mao, Jongyoon Han)
Sieving in Periodic Free-Energy Landscapes Created by
Patterned Nanofilter Arrays. (Jianping Fu, Juhwan Yoo,
Nanofluidic Filter Array for High-Throuput Biomolecule
Separation. (Pan Mao, Jongyoon Han)
of Effect of Structural Parameters of the Nanofilter
Array on Biomolecule Separation. Dispersion of Fractionated
Biomolecule Bands in the Nanofilter Array. (Hansen Bow,
Jianping Fu, Craig Rothman, Jongyoon Han)
Design and Fabrication of Anisotropic Two-Dimensional Nanofilter Array (Anisotropic Nanofilter Array: ANA) for Continuous-Flow
Biomolecule Separation. (Jianping Fu, Jongyoon
- Continuous-Flow Ogston Sieving-Based Separation of Short DNA and SDS-Protein Complexes and Entropic Trapping-Based Separation of Long DNA Through the ANA. (Jianping Fu, Jongyoon Han)
- Continuous-Flow Size- or Charge-Based Separation of Proteins under Native Conditions through the ANA. (Reto B. Schoch, Jianping Fu, Jongyoon Han)
- Suppressing air
bubbles trapped in the one-dimensional nanofilter array. (Real time. Air bubbles trapped in the
nanofilter deep regions were suppressed by EOF flow. Video by Jianping Fu.)
- Fast SDS-protein complex
separation in one-dimensional periodic array of nanofilter. (10X speed. Three proteins separated
within 30 sec, Eav = 100 V/cm. Video by Jianping Fu.)
separation of short DNA (the PCR marker, 50 - 1,000 bp) through two-dimensional periodic array
of nanofilters (Anisotropic Nanofilter Array: ANA), based
on the Ogston sieving mechanism. (This video was taken with exposure time of 1300 ms/image and image size of 1300 µm X 1620 µm. The time scale in this movie has been compressed by a factor of 20. At the beginning of the movie, only Ey = 25 V/cm was applied. The orthogonal field Ex = 35 V/cm was applied at 3 sec in the movie, and the separation was finished at about 12 sec in the movie. Video by Jianping Fu.)
separation of long DNA (the Lambda DNA-Hind III digest, 2,000 - 23,000 bp) through ANA, based on
the entropic trapping mechanism. (This video was taken with exposure time of 600 ms/image and image size of 3270 µm X 4080 µm. The time scale in this movie has been compressed by a factor of 20. At the beginning of the movie, only Ey = 100 V/cm was applied. The orthogonal field Ex = 185 V/cm was applied at 1 sec in the movie, and the separation was finished at about 5 sec in the movie. Video by Jianping Fu.)
separation of SDS-protein complexes through ANA. (This video was taken with exposure time of 1000 ms/image and image size of 3270 µm X 4080 µm. The time scale in this movie has been compressed by a factor of 20. At the beginning of the movie, only the vertical field Ey = 50 V/cm was applied. The horizontal field Ex = 75 V/cm was applied at 1 sec in the movie, and the separation was finished at about 6 sec in the movie. Video by Jianping Fu.)
- Schoch, R. B., Fu, J., Bow, H. & Han, J. MicroScale Bioseparation 2007 Symposium, Vancouver, Canada.
- Fu, J.*, Schoch, R. B.*, Stevens, A. L., Tannenbaum, S. R. & Han, J. Nature Nanotech. 2, 121-128 (2007). (pdf)
- Bow, H., Fu, J., Rothman, C. & Han, J. Anal. Chem., submitted (2006).
- Fu, J. & Han, J. Proceedings of the MicroTAS 2006 Symposium, Tokyo, Japan, vol. 1, pp. 519-521. (pdf)
- Fu, J., Yoo, J. & Han, J. Phys. Rev. Lett.
97, 018103.1-3 (2006). (pdf‡)
Fu, J., Mao, P. & Han, J. Appl. Phys.
Lett. 87, 263902.1-3 (2005). (pdf†)
Fu, J. & Han, J. American Physical Society
National March Meeting 2006, Baltimore, Maryland. (pdf)
- Fu, J. & Han, J. Proceedings of the MicroTAS 2005 Symposium, Boston, MA, vol. 2, pp. 1531-1533. (pdf)
- Fu, J. & Han, J. 2005 Gordon Research Conf. on the Physics and Chemistry of Microfluidics, Oxford, UK.
- Fu, J. & Han, J. MicroScale Bioseparation 2005 Symposium, New Orleans, Louisiana.
- Fu, J. & Han, J. Proceedings of the MicroTAS 2004 Symposium, Malmo, Sweden, vol. 1, pp. 285-287.
* These authors contributed equally to this work.
† Copyright by American Institute of Physics. This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics.
‡ Copyright by the American Physical Society.