DOI QR코드

DOI QR Code

고효율 바이오물질 분리 및 농축을 위한 나노필터소자제작

Fabrication of Nano-filter Device for High Efficient Separation and Concentration of Biomolecules

  • 허윤석 (한국기초과학지원연구원 물성과학연구부) ;
  • 최봉길 (한국과학기술원 생명화학공학과) ;
  • 홍원희 (한국과학기술원 생명화학공학과)
  • Huh, Yun Suk (Division of Material Science, Korea Basic Science Institute) ;
  • Choi, Bong Gill (Department of Chemical & Biomolecular Engineering (BK21 program), KAIST) ;
  • Hong, Won Hi (Department of Chemical & Biomolecular Engineering (BK21 program), KAIST)
  • 투고 : 2012.02.09
  • 심사 : 2012.03.28
  • 발행 : 2012.08.01

초록

본 연구에서는 알루미나 나노 템플레이트(anodic alumina oxide; AAO)를 이용하여 신속하면서도 효과적으로 나노입자 및 바이오물질을 분리, 농축할 수 있는 나노필터 소자를 개발하였다. 본 연구에서 사용한 나노필터 소자는 유체의 주입 및 흐름이 가능한 미세유체채널(microfluidic channel)을 PDMS에 패터닝하였다. 위아래로 형성된 PDMS 미세유체채널 사이로, 다양한 크기의 나노 다공을 형성하고 있는 AAO 막을 삽입하여 크기에 따른 나노입자 및 바이오 물질을 분리할 수 있었다. 위아래로 PDMS 유체채널과 AAO 분리막을 집적하고, 최종적으로 아크릴레이트 플락스틱(acrylic plastic)으로 전체 소자를 고정하여 나노필터유체소자를 제작하였다. 완성된 나노필터소자를 이용하여 나노입자의 농축효율 및 은나노입자가 뭉쳐져있는 필터존(filtration zone)으로부터 뎅기 바이러스(dengue virus)를 표면증강라만(surface enhanced Raman scattering)분석법에 의해 검출할 수 있었다.

Here, we develop a new nanofilter device for the rapid and efficient separation of nanoparticles and biomolecules, exploiting the use of AAO mebrane with ordered nanopores in the range from 20 nm to 200 nm. Briefly, the chip comprises of a series of the upper and lower PDMS channels containing embedded inlet and outlet ports, and $50{\mu}m$ width microfluidic channel, and AAO membrane to be made the filtering zone. After assembling these components, the acrylate plastic plates were used to fix the device on the top and bottom side. When introducing the samples into the inlet ports of the upper PDMS channel, we were able to separate and concentrate the nanoparticles and target molecules at the filtering zone, and to elute the solutions containing the unwanted materials toward the lower PDMS channels normal to the direction of AAO membrane. To demonstrate the usefulness of the device we apply it to the SERS detection of nucleic acid sequences associated with Dengue virus serotype 2. We report a limit of detection for Dengue sequences of 300 nM and show excellent enhancement of Raman signals from the filter zone of the nanofilter device.

키워드

과제정보

연구 과제 주관 기관 : 한국기초과학지원연구원

참고문헌

  1. Deen, W. M., "Hindered Transport of Large Molecules in Liquid- filled Pores," AIChE J., 33, 1409-1425(1987). https://doi.org/10.1002/aic.690330902
  2. Nishizawa, M., Menon, V. P. and Martin, C. R., "Metal Nanotubule Membranes with Electrochemically Switchable Ion-Transport Selectivity," Science, 268, 700-702(1995). https://doi.org/10.1126/science.268.5211.700
  3. Jirage, K. B., Hulteen, J. C. and Martin, C. R., "Nanotubule-Based Molecular-Filtration Membranes," Science, 278, 655-658(1997). https://doi.org/10.1126/science.278.5338.655
  4. Che, G., Lakshmi, B. B., Fisher, E. R. and Martin, C. R., "Carbon Nanotubule Membranes for Electrochemical Energy Storage and Production," Nature, 393, 346-349(1998). https://doi.org/10.1038/30694
  5. Lee, S. B., Mitchell, D. T., Trofin, L., Nevanen, T. K., Soderlund, H. and Martin, C. R., "Antibody-Based Bio-Nanotube Membranes for Enantiomeric Drug Separations," Science, 296, 2198-2200(2002). https://doi.org/10.1126/science.1071396
  6. Fu, J., Mao, P. and Han, J., "Nanofilter Array Chip for Fast Gelfree Biomolecule Separation," Appl. Phys. Lett., 87, 263902-263902-3(2005). https://doi.org/10.1063/1.2149979
  7. Fu, J., Yoo, J. and Han, J., "Molecular Sieving in Periodic Free- Energy Landscapes Created by Patterned Nanofilter Arrays," Phys. Rev. Lett., 97, 018103-018106(2006). https://doi.org/10.1103/PhysRevLett.97.018103
  8. Park, S., Huh, Y. S., Craighead, H. G. and Erickson, D., "A Method for Nanofluidic Device Prototyping Using Elastomeric Collapse," Proc. Natl. Acad. Sci. USA, 106, 15549-15554(2009). https://doi.org/10.1073/pnas.0904004106
  9. Huh, Y. S., Choi, J. H., Park, T. J., Hong, Y. K., Hong, W. H. and Lee, S. Y., "Microfluidic Cell Disruption System Employing a Magnetically Actuated Diaphragm," Electrophoresis, 28, 4748-4757(2007). https://doi.org/10.1002/elps.200700366
  10. Lee, E. Z., Huh, Y. S., Jun, Y. S., Won, H. J., Hong, Y. K., Park, T. J., Lee, S. Y. and Hong, W. H., "Removal of Bovine Serum Albumin Using Solid-phase Extraction with In-situ Polymerized Stationary Phase in a Microfluidic Device," J. Chromato. A., 1187, 11-17(2008). https://doi.org/10.1016/j.chroma.2008.01.084
  11. Zaytseva, N. V., Montagna, R. A., Lee, E. M. and Baeumner, A. J., "Multi-analyte Single-membrane Biosensor for the Serotypespecific Detection of Dengue Virus," Anal. Bioanal. Chem., 380, 46-53(2004).
  12. Franz, A. W. E., Sanchez-Vargas, I., Adelman, Z. N., Blair, C. D., Beaty, B. J., James, A. A. and Olson, K. E., "Engineering RNA Interference-based Resistance to Dengue Virus type 2 in Genetically Modified Aedes Aegypti," Proc. Natl. Acad. Sci. USA, 103, 4198-4203(2006). https://doi.org/10.1073/pnas.0600479103
  13. Lowe, A. J., Huh, Y. S., Strickland, A. D., Erickson, D. and Batt, C. A., "Multiplex Single Nucleotide Polymorphism Genotyping Utilizing Ligase Detection Reaction Coupled Surface Enhanced Raman Spectroscopy," Anal. Chem., 82, 5810-5814(2010). https://doi.org/10.1021/ac100921b
  14. Brown, R., Smith, W. E. and Graham, D., "Synthesis of a Ben-zotriazole Phosphoramidite for Attachment of Oligonucleotides to Metal Surfaces," Terahedron Lett., 42, 2197-2200(2001). https://doi.org/10.1016/S0040-4039(01)00108-3
  15. Cao, Y. C., Jin, R. and Mirkin, C. A., "Nanoparticles with Raman Spectroscopic Fingerprints for DNA and RNA Detection," Science, 297, 1536-1540(2002). https://doi.org/10.1126/science.297.5586.1536
  16. Huh, Y. S., Chung, A. J., Cordovez, B. and Erickson, D., "Enhanced on-chip SERS Based Biomolecular Detection Using Electrokinetically Active Microwells," Lab Chip, 9, 433-439(2009). https://doi.org/10.1039/b809702j
  17. Abu-Hatab, N. A., John, J. F., Oran, J. M. and Sepaniak, M. J., "Multiplexed Microfluidic Surface-Enhanced Raman Spectroscopy," Appl. Spectrosc., 61, 1116-1122(2007). https://doi.org/10.1366/000370207782217842

피인용 문헌

  1. 인플루엔자 바이러스 검출을 위한 종이 기반 neuraminidase 효소 활성 평가 센서 개발 vol.54, pp.3, 2012, https://doi.org/10.9713/kcer.2016.54.3.380