TITLE

FRET detection of Octamer-4 on a protein nanoarray made by size-dependent self-assembly

AUTHOR(S)
Tran, Phat L.; Gamboa, Jessica R.; You, David J.; Jeong-Yeol Yoon
PUB. DATE
September 2010
SOURCE
Analytical & Bioanalytical Chemistry;Sep2010, Vol. 398 Issue 2, p759
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
n alternative approach for fabricating a protein array at nanoscale is suggested with a capability of characterization and/or localization of multiple components on a nanoarray. Fluorescent micro- and nanobeads each conjugated with different antibodies are assembled by size-dependent self-assembly (SDSA) onto nanometer wells that were created on a polymethyl methacrylate (PMMA) substrate by electron beam lithography (EBL). Antibody-conjugated beads of different diameters are added serially and electrostatically attached to corresponding wells through electrostatic attraction between the charged beads (confirmed by zeta potential analysis) and exposed p-doped silicon substrate underneath the PMMA layer. This SDSA method is enhanced by vibrated-wire-guide manipulation of droplets on the PMMA surface containing nanometer wells. Saturation rates of antibody-conjugated beads to the nanometer patterns are up to 97% under one component and 58–70% under two components nanoarrays. High-density arrays (up to 40,000 wells) could be fabricated, which can also be multi-component. Target detection utilizes fluorescence resonance energy transfer (FRET) from fluorescent beads to fluorescent-tagged secondary antibodies to Octamer-4 (Oct4), which eliminates the need for multiple steps of rinsing. The 100 nm green beads are covalently conjugated with anti-Oct4 to capture Oct4 peptides (39 kDa); where the secondary anti-Oct4 and F(ab) fragment of anti-gIgG tagged with phycoerythrin are then added to function as an indicator of Oct4 detection. FRET signals are detected through confocal microscopes, and further confirmed by Fluorolog3 spectrofluorometer. The success rates of detecting Oct4 are 32% and 14% of the beads in right place under one and two component nanoarrays, respectively. Ratiometric FRET is used to quantify the amount of Oct4 peptides per each bead, which is estimated about 2 molecules per bead.
ACCESSION #
53556223

 

Related Articles

  • Hybrid template approach achieves sub-30 nm NIL.  // Laser Focus World;Nov2008, Vol. 44 Issue 11, p17 

    The article describes a hybrid molecular-ruler approach developed at the Pennsylvania State University which achieves sub-30 nm patterning over a full water. It shows how electron-beam lithography or other methods create the first level of metallic features. A final etching process transfers the...

  • Around the bend. Heber, Joerg // Nature Materials;Dec2011, Vol. 10 Issue 12, p902 

    The article presents a study which aims to develop a deposition method that enables the fabrication of nanoscale structures on flexible substrates, which was based on nanostencil lithography.

  • Patterning concept for sculptured nanostructures with arbitrary periods. Patzig, C.; Zajadacz, J.; Zimmer, K.; Fechner, R.; Khare, C.; Rauschenbach, B. // Applied Physics Letters;9/7/2009, Vol. 95 Issue 10, p103107 

    An approach to deposit sculptured nanostructures with arbitrary interstructure spacing is presented. Based on a combination of glancing angle deposition with a preceding two-step electron beam lithography substrate patterning, the concept allows the deposition of nanostructures on artificial...

  • Nanoimprint lithography of plasmonic platforms for SERS applications. Barcelo, Steven; Wu, Wei; Li, Xuema; Li, Zhiyong; Williams, R. // Applied Physics A: Materials Science & Processing;Nov2015, Vol. 121 Issue 2, p443 

    In this review, we describe the use of nanoimprint lithography in our group to fabricate plasmonic platforms with nanometer-scale critical features that would be significantly more expensive using other fabrication techniques: 3-D cones that have tips with a sub-10 nm radius of curvature, active...

  • Advanced Monitoring for Nanoimprint Lithography. Hocheng, H.; Hsu, W. H. // AIP Conference Proceedings;1/17/2011, Vol. 1315 Issue 1, p1237 

    Nanoimprint lithography has the advantages of high throughput, sub-10-nm feature and low cost. A problem of incomplete filling rate is, however, encountered in the imprinting process and needs to be understood for pattern fidelity. The filling rate is often measured with a scanning electron...

  • Biophysics: Nanocombinatorics for biology.  // Nature Methods;May2012, Vol. 9 Issue 5, p436 

    The article focuses on the study conducted by L. T. Giam and colleagues which describes the methods to study cell-substrate interactions using massively parallel polymer pen lithography to generate substrates with customized patterns.

  • Photothermal actuation in nanomechanical waveguide devices. Pernice, Wolfram H. P.; Li, Mo; Tang, Hong X. // Journal of Applied Physics;Jan2009, Vol. 105 Issue 1, p014508 

    We analyze the photothermal response of nanoelectromechanical systems (NEMS) integrated in a silicon photonic circuit over a wide frequency range. The dynamic response of NEMS devices is studied using a two-color pump-probe scheme in an on-chip photonic Mach–Zehnder interferometer. The...

  • Interfacial structure and chemistry of epitaxial CoFe2O4 thin films on SrTiO3 and MgO substrates. Xie, S.; Cheng, J.; Wessels, B. W.; Dravid, V. P. // Applied Physics Letters;11/3/2008, Vol. 93 Issue 18, p181901 

    Interfacial structure and chemistry of epitaxial CoFe2O4 thin films on (001)_SrTiO3 and (001)_MgO substrates were investigated using high-angle-annular-dark-field imaging combined with electron-energy-loss-spectra (EELSs) analyses at a subnanometer scale. The results show that CoFe2O4/SrTiO3...

  • Shift of isoelectric point in extended nanospace investigated by streaming current measurement. Morikawa, Kyojiro; Mawatari, Kazuma; Kazoe, Yutaka; Tsukahara, Takehiko; Kitamori, Takehiko // Applied Physics Letters;9/19/2011, Vol. 99 Issue 12, p123115 

    Isoelectric points in extended nanochannels (580-2720 nm) fabricated on fused-silica substrates were measured using the streaming current method. The isoelectric point obtained in a 2720 nm channel was almost the same as the isoelectric point reported for the bulk (2.6-3.2). However, the...

Share

Read the Article

Courtesy of VIRGINIA BEACH PUBLIC LIBRARY AND SYSTEM

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics