Direct Write Protein Patterns for Multiplexed Cytokine Detection from Live Cells Using Electron Beam Lithography

Date

2016-01-01

Author

Lau, Uland Y.
Saxer, Sina S.
Lee, Juneyoung
Bat, Erhan
Maynard, Heather D.

Metadata

Show full item record

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Item Usage Stats

226
views

0
downloads

Simultaneous detection of multiple biomarkers, such as extracellular signaling molecules, is a critical aspect in disease profiling and diagnostics. Precise positioning of antibodies on surfaces, especially at the micro- and nanoscale, is important for the improvement of assays, biosensors, and diagnostics on the molecular level, and therefore, the pursuit of device miniaturization for parallel, fast, low-volume assays is a continuing challenge. Here, we describe a multiplexed cytokine immunoassay utilizing electron beam. lithography and a trehalose glycopolymer as a resist for the direct writing of antibodies on silicon substrates, allowing for micro- and nanoscale precision of protein immobilization. Specifically, anti-interleukin 6 (IL-6) and antitumor necrosis factor alpha (TNF alpha) antibodies were directly patterned. Retention of the specific binding properties of the patterned antibodies was shown by the capture of secreted cytokines from stimulated RAW 264.7 macrophages. A sandwich immunoassay was employed using gold nanoparticles and enhancement with silver for the detection and visualization of bound tytokines to the patterns by localized surface plasmon resonance detected with dark-field microscopy. Multiplexing with both IL-6 and TNF alpha on, a single chip was also successfully demonstrated with high specificity and in relevant cell culture conditions and at different times after cell stimulation. The direct fabrication of capture antibody patterns for cytokine detection described here could be useful for biosensing applications.

Subject Keywords

General Engineering, General Physics and Astronomy, General Materials Science

URI

https://hdl.handle.net/11511/48584

Journal

ACS NANO

DOI

https://doi.org/10.1021/acsnano.5b05781

Collections

Department of Chemical Engineering, Article

Suggestions

OpenMETU
Core

Integration of a Genetically Encoded Calcium Molecular Sensor into Photopolymerizable Hydrogels for Micro-Optrode-Based Sensing Kahyaoğlu, Leyla Nesrin; Park, Joon Hyeong; Rickus, Jenna L. (American Chemical Society (ACS), 2017-09-20) Genetically encoded molecular-protein sensors (GEMS) are engineered to sense and quantify a wide range of biological substances and events in cells, in vitro and even in vivo with high spatial and temporal resolution. Here, we aim to stably incorporate these proteins into a photopatternable matrix, while preserving their functionality, to extend the application of these proteins as spatially addressable optical biosensors. For this reason, we examined the fabrication of 3D hydrogel microtips doped with a ge...
Computational modeling of coupled cardiac electromechanics incorporating cardiac dysfunctions Berberoglu, Ezgi; Solmaz, H. Onur; Göktepe, Serdar (Elsevier BV, 2014-11-01) Computational models have huge potential to improve our understanding of the coupled biological, electrical, and mechanical underpinning mechanisms of cardiac function and diseases. This contribution is concerned with the computational modeling of different cardiac dysfunctions related to the excitation-contraction coupling in the heart. To this end, the coupled problem of cardiac electromechanics is formulated through the conservation of linear momentum equation and the excitation equation formulated in th...
Numerical study on effects of computational domain length on flow field in standing wave thermoacoustic couple MERGEN, SÜHAN; Yıldırım, Ender; TÜRKOĞLU, HAŞMET (Elsevier BV, 2019-03-01) For the analysis of thermoacoustic (TA) devices, computational methods are commonly used. In the computational studies found in the literature, the flow domain has been modelled differently by different researchers. A common approach in modelling the flow domain is to truncate the computational domain around the stack, instead of modelling the whole resonator to save computational time. However, where to truncate the domain is not clear. In this study, we have investigated how the simulation results are aff...
Integrability of a generalized Ito system: The Painleve test Karasu-Kalkanli, A; Karasu, Atalay; Sakovich, SY (Physical Society of Japan, 2001-05-01) It is shown that a generalized Ito system of four coupled nonlinear evolution equations passes the Painleve test for integrability in five distinct cases, two of which were introduced recently by Tam et al. [H. W. Tam, X. B. Hu, D. L. Wang: J. Phys. Sec. Jpn. 68 (1999) 369]. A conjecture is formulated on integrability of a vector generalization of the Ito system.
RECONSTRUCTION OF PERMITTIVITY AND CONDUCTIVITY PROFILES OF A DIELECTRIC SLAB IN THE TIME DOMAIN BY DESCENT METHODS ONDER, M; Kuzuoğlu, Mustafa (Institution of Engineering and Technology (IET), 1992-10-01) An optimisation approach is presented for the problem of reconstructing the permittivity and conductivity profiles of a dielectric slab from the reflected and transmitted field data. The problem is treated as an optimal control problem where the norm of the difference of measured and calculated boundary data is minimised subject to the state equation governing the system. The original constrained optimisation problem is reduced to the evaluation of stationary points of an augmented functional which is obtai...

Citation Formats

U. Y. Lau, S. S. Saxer, J. Lee, E. Bat, and H. D. Maynard, “Direct Write Protein Patterns for Multiplexed Cytokine Detection from Live Cells Using Electron Beam Lithography,” ACS NANO, pp. 723–729, 2016, Accessed: 00, 2020. [Online]. Available: https://hdl.handle.net/11511/48584.