Award Abstract # 1201878
Molecularly Engineered Artificial Nanopores with Differential Selectivity and Sensitivity

NSF Org: ECCS
Division of Electrical, Communications and Cyber Systems
Recipient: UNIVERSITY OF TEXAS AT ARLINGTON
Initial Amendment Date: March 4, 2012
Latest Amendment Date: April 22, 2014
Award Number: 1201878
Award Instrument: Standard Grant
Program Manager: Usha Varshney
ECCS
 Division of Electrical, Communications and Cyber Systems
ENG
 Directorate for Engineering
Start Date: March 15, 2012
End Date: February 29, 2016 (Estimated)
Total Intended Award Amount: $360,000.00
Total Awarded Amount to Date: $388,000.00
Funds Obligated to Date: FY 2012 = $360,000.00
FY 2013 = $12,000.00

FY 2014 = $16,000.00
History of Investigator:
  • Samir Iqbal (Principal Investigator)
    smiqbal@utrgv.edu
  • Richard Timmons (Co-Principal Investigator)
  • Purnendu Dasgupta (Co-Principal Investigator)
Recipient Sponsored Research Office: University of Texas at Arlington
701 S NEDDERMAN DR
ARLINGTON
TX  US  76019-9800
(817)272-2105
Sponsor Congressional District: 25
Primary Place of Performance: University of Texas at Arlington
TX  US  76019-0145
Primary Place of Performance
Congressional District:
25
Unique Entity Identifier (UEI): LMLUKUPJJ9N3
Parent UEI:
NSF Program(s): EPMD-ElectrnPhoton&MagnDevices
Primary Program Source: 01001213DB NSF RESEARCH & RELATED ACTIVIT
01001314DB NSF RESEARCH & RELATED ACTIVIT

01001415DB NSF RESEARCH & RELATED ACTIVIT
Program Reference Code(s): 108E, 103E, 9102, 8028, 9251
Program Element Code(s): 151700
Award Agency Code: 4900
Fund Agency Code: 4900
Assistance Listing Number(s): 47.041

ABSTRACT

Objectives:

This proposal addresses a number of fundamental and important obstacles that have limited the utility of nanopore sensing technology. Specifically, nanopore size, surface composition and stability of the pores are addressed to broaden this technology to include important new areas, e.g. sequencing of genomes, as but one of many examples. To help expedite progress in this area, our objective involves fabrication and thorough evaluation of a new approach to nanopore sensor technology. Specifically, we propose construction of artificial pores in which both the size and surface chemistry of the pores are precisely controlled over a wide range of pore sizes. For this purpose, a novel combination of thermal processing, followed by pulsed plasma chemical vapor deposition, will be used to shrink pore sizes controllably and reproducibly, while simultaneously varying surface chemistry. Availability of these molecularly engineered nanopores will hopefully overcome limitations currently encountered in analytical applications of this technology. An additional important objective involves extension of this technology to several new, high profile applications to be made available via our approach, e.g., selective differential detection by two oppositely chirally functionalized nanopores which will revolutionize chromatographic measurement of chiral compounds; similar schemes will be possible in many other cases.

Intellectual Merits of the Proposed Activity?

To date, nanopore fabrication has focused primarily on low-throughput serial approaches, with little use of bottom-up technology. In general, key issues such as stability of nanopore/fluid interfaces, reproducibility of nanopore surface properties, extended pore stability and the need for robust chemical functionalization of the nanopores remain. The ready availability of mechanically stable nanopores, having a range of well-defined diameters and surface chemistries, as described in this proposal, represents a transformative advance in this area. New insights will be gained from selective interactions in nanopores, providing quantitative handles to evaluate the molecular responses of ligands and to define novel, chemical means of interrogating targeted analytes. This will stimulate additional studies: control of translocation times of analytes through the pores (an immensely important present problem) and the use of chemistry and size differentiated nanopore arrays. This innovation will also help overcome and replace the present labor-intensive and low-throughput fabrication methods.

Broader Impacts of the Proposed Activity?

The proposed project will impact many areas that depend on the confluence of sciences and engineering. Examples include design of bio-inspired systems, sensors for environment and living systems, etc. The basic principles involved can be integrated into all levels of education. Graduate and undergraduate (UG) students will be engaged and introduced to exciting new dimensions of analytical chemistry, biochemistry and solid-state fabrication through development of a cross-listed course module on the bio-nano interface. The research outcomes will be also used to develop integrative participatory modules at our presently conducted Summer Camps (for middle/high-school students) and Girlgeneering Camps (for female high school students) to attract future adults to STEM careers. These outreach endeavors will be pursued: (1) Seminars/lab-tours for involvement and retention of UGs in research; (2) Engaging minority students through the McNair Fellows program; (3) Dynamic Facebook presence for the projection/exposure/discussion of the research; (4) Engagement of K-12 students and teachers through live webcasts. The results of the proposed research and education endeavors will be disseminated not only through peer-reviewed articles and conferences, but also through public media (radio, newspaper, weblogs, public displays). UTA Chemistry participates in the local State Fair. UTA has the largest digital planetarium in the Metroplex, with extremely heavy K-12 traffic. We plan to develop a small clip on nanopore sensors.

PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

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A. Hafeez, W. Asghar, M. M. Rafique, S. M. Iqbal and A. R. Butt "GPU-based real-time detection and analysis of biological targets using solid-state nanopores" Medical & Biological Engineering & Computing , v.50 , 2012 , p.605-615
J. A. Billo, J. Jones, W. Asghar, R. L. Carter and S. M. Iqbal "Viscosity and Surface-Free Energy Effects in Thermal Shrinking of Solid-State Nanopores" Applied Physics Letters , v.100 , 2012 , p.233107
M. A. I. Mahmood, A. Adnan and S. M. Iqbal "3D Structural Integrity and Interactions of Single-stranded Protein-binding DNA in a Functionalized Nanopore," Journal of Physical Chemistry B , v.118 , 2014 , p.5799
M. A. I. Mahmood, W. Ali, A. Adnan and S. M. Iqbal "3D Structural Integrity and Interactions of Single-stranded Protein-binding DNA in a Functionalized Nanopore" The Journal of Physical Chemistry B , v.118 , 2014 , p.5799?5806 10.1021/jp411820w
M. R. Hasan, M. A. I. Mahmood, A. Ashfaq and S. M. Iqbal "Electrostatic Denaturation during Solid-State Nanopore Translocating" Modeling of Biomolecular Systems Interactions, Dynamics, and Allostery: Bridging Experiments and Computations Meeting , 2014

PROJECT OUTCOMES REPORT

Disclaimer

This Project Outcomes Report for the General Public is displayed verbatim as submitted by the Principal Investigator (PI) for this award. Any opinions, findings, and conclusions or recommendations expressed in this Report are those of the PI and do not necessarily reflect the views of the National Science Foundation; NSF has not approved or endorsed its content.

This team developed solid-state nanopores as single-molecule sensors for detection and enumeration of a cancer biomarker called EGFR.  The point-of-care (POC) detection of cancer and other disease biomarkers is a pressing need. Devices for POC must be ultrasensitive, fast, accurate, low priced and should be easy to use.

The solid-state nanopores provided single molecule detection based on the resistive-pulse enumeration. When a molecule hindered the ionic flow through the nanopore, it was registered as a unique electrical pulse in the baseline ionic current trace. Analysis of these electrical pulses has been done to determine size and charge of molecules, length of nucleic acids, protein size, folding state and molecular agglomeration. 

To keep the measurement process simple, instead of using a functionalized nanopore, a bare nanopore was used and in-solution binding of EGFR with anti-EGFR aptamer was used to impart selectivity. Anti-EGFR aptamer has very high affinity for EGFR and it is very selective as well. Aptamer binding to the protein altered the overall charge and mass of the complex as compared to the unbounded EGFR. This change was readily identified from the analysis of registered pulses. The single molecule sensitivity gave us capability to detect and enumerate each molecule from the sample. 

The experimental work was done in cnjunction with the theoretical modelling. The stability of DNA structure under applied electric field, the mechanism of protein translocation through DNA functionalized nanopore and the travel of protein-DNA complex through a bare nanopore were first studied using all-atom molecular dynamic simulations. The simulations guided the design of the experiments.  

 

 


Last Modified: 05/29/2016
Modified by: Samir M Iqbal

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