Protein nanoarray made by size-dependent self-assembly for detection of mouse immunoglobulin G and octamer-4

Phat L. Tran, Yee Tchao, David J. You, Jeong-Yeol Yoon

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

An alternative approach for fabricating a protein array at nanoscale (<100 nm) is suggested with a capability of characterization and/or localization of multiple components on a nanoarray. Basically, fluorescent micro- and nanospheres each conjugated with different proteins are size-dependently self-assembled (SDSA) onto these nanometer wells that were created on the polymethyl methacrylate (PMMA) substrate by electron beam lithography (EBL). Particles of different diameters are added serially, and electrostatically attached to the corresponding wells through electrostatic attraction between the carboxylic groups of the spheres and p-doped silicon substrate underneath the PMMA layer. This SDSA was enhanced by wire-guide manipulation of droplets on the surface containing nanometer wells. Target detection utilizes fluorescence resonance energy transfer (FRET) from fluorescent beads to target (mouse immunoglobulin G = mIgG or Octamer-4 = Oct4) and its antibody bound on the beads. The 180 nm blue beads are conjugated with mIgG to capture anti-mIgG-FITC. The 50 nm green and 100 nm yellow-green beads are conjugated with anti-Oct4 to capture Oct4 peptides; where the secondary anti-Oct4 tagged with phycoerythrin via F(ab)2 fragment is then added to function as an indicator of Oct4 detection. These protein-conjugated particles are added serially from the largest to the smallest and the particles are successfully self-assembled to the respective nanometer wells to achieve sizedependent self-assembly. FRET signals are detected through fluorescence and confocal microscopes, and further confirmed by Fluorolog3 spectrofluorometer. Therefore, SDSA is a valuable approach for the fabrication of multiple components array; and FRET is a useful biorecognition technique for the detection of mIgG, Oct4 or other targets of interest.

Original languageEnglish (US)
Title of host publicationProgress in Biomedical Optics and Imaging - Proceedings of SPIE
Volume7313
DOIs
StatePublished - 2009
EventSmart Biomedical and Physiological Sensor Technology VI - Orlando, FL, United States
Duration: Apr 16 2009Apr 17 2009

Other

OtherSmart Biomedical and Physiological Sensor Technology VI
CountryUnited States
CityOrlando, FL
Period4/16/094/17/09

Fingerprint

Fluorescence Resonance Energy Transfer
beads
Self assembly
mice
self assembly
resonance fluorescence
Immunoglobulin G
Polymethyl Methacrylate
proteins
Proteins
Polymethyl methacrylates
energy transfer
polymethyl methacrylate
Phycoerythrin
Nanospheres
Protein Array Analysis
Electron beam lithography
Fluorescein-5-isothiocyanate
Silicon
Substrates

Keywords

  • E-Beam lithography
  • Fluorescence resonance energy transfer (FRET)
  • Nanometer pattern generation system
  • Wire-guide droplet manipulation

ASJC Scopus subject areas

  • Atomic and Molecular Physics, and Optics
  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Radiology Nuclear Medicine and imaging

Cite this

Tran, P. L., Tchao, Y., You, D. J., & Yoon, J-Y. (2009). Protein nanoarray made by size-dependent self-assembly for detection of mouse immunoglobulin G and octamer-4. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE (Vol. 7313). [731306] https://doi.org/10.1117/12.817888

Protein nanoarray made by size-dependent self-assembly for detection of mouse immunoglobulin G and octamer-4. / Tran, Phat L.; Tchao, Yee; You, David J.; Yoon, Jeong-Yeol.

Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7313 2009. 731306.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Tran, PL, Tchao, Y, You, DJ & Yoon, J-Y 2009, Protein nanoarray made by size-dependent self-assembly for detection of mouse immunoglobulin G and octamer-4. in Progress in Biomedical Optics and Imaging - Proceedings of SPIE. vol. 7313, 731306, Smart Biomedical and Physiological Sensor Technology VI, Orlando, FL, United States, 4/16/09. https://doi.org/10.1117/12.817888
Tran PL, Tchao Y, You DJ, Yoon J-Y. Protein nanoarray made by size-dependent self-assembly for detection of mouse immunoglobulin G and octamer-4. In Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7313. 2009. 731306 https://doi.org/10.1117/12.817888
Tran, Phat L. ; Tchao, Yee ; You, David J. ; Yoon, Jeong-Yeol. / Protein nanoarray made by size-dependent self-assembly for detection of mouse immunoglobulin G and octamer-4. Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7313 2009.
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