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Handheld Device for Rapid In-Field Assay of Listeria Monocytogenes Using Designed Supercharged Binding Proteins for Enhanced Surface Plasmon Resonance (SPR)

Award Information
Agency: Department of Health and Human Services
Branch: Food and Drug Administration
Contract: 1R43FD005693-01
Agency Tracking Number: R43FD005693
Amount: $77,807.00
Phase: Phase I
Program: SBIR
Solicitation Topic Code: FDA
Solicitation Number: PA15-269
Timeline
Solicitation Year: 2015
Award Year: 2016
Award Start Date (Proposal Award Date): 2016-09-15
Award End Date (Contract End Date): 2017-12-31
Small Business Information
419 LAFAYETTE ST
New York, NY 10003-7033
United States
DUNS: 145785528
HUBZone Owned: No
Woman Owned: No
Socially and Economically Disadvantaged: No
Principal Investigator
 LORI LEPAK
 (718) 484-7033
 navallone@phoebus-opto.com
Business Contact
 NICOLE AVALLONE
Phone: (718) 484-7033
Email: navallone@phoebus-opto.com
Research Institution
N/A
Abstract

Project Abstract
The proposed project aims to create a new weapon in the arsenal of the CDC for use in the `winnable battleandapos
for improved food safety a low cost hand held high sensitivity rapid sensing platform to detect pathogens
and biological and chemical toxins in the field such as on farms at food processing plants and in medical
facilities This sensor device incorporates a new class of supercharged binding proteins created through
newly developed biodesign technologies onto a sensor chip with a metastructured gold film to produce a one
thousandfold enhancement in the surface plasmon resonance SPR shift upon binding of the target pathogen
Other than the engineered binding protein the overall device uses only low cost disposable detector chips
fabricated via standard semiconductor fabrication processes and off the shelf optical components arranged in
a more compact and robust geometry than any other known commercially available SPR detector
This research will initially target the detection of Listeria monocytogenes serotype b which is responsible for
an estimated deaths and $ billion in outbreak containment costs each year The broader underlying
principles can be applied to tap into the rapidly growing In Vitro Diagnostics IVD market valued globally at
$ billion in to achieve the long term goal of the rapid sensitive highly specific detection of most viral
and gram negative bacterial pathogens which are relevant to public health both domestically and abroad
The proposed research will proceed with specific aims Design and optimize short chain variable fragment
ScFv proteins based upon wild type monoclonal antibodies to L monocytogenes serotype b the target
pathogen Supercharge the designed ScFv protein by mutating antibody residues not essential for target
binding Use standard semiconductor fabrication techniques to fabricate a gold plasmonic metasurface on a
transparent fused silica substrate and attach the designed ScFv protein to it using standard gold thiol and
biotin streptavidin chemistries and Measure the SPR resonance shift as ScFv binds to its target ie Listeria
As a proof of concept of the highest risk highest reward component of the device i e development of the
supercharged binding protein a model heme binding protein named Mega was designed synthesized and
characterized Mega demonstrated a large binding induced conformational change which caused a
refractive index change which is calculated to correspond to a fold enhancement in SPR signal
relative to commercially available ligand antibody detection schemes Structural elements of Mega were
shown to undergo a phase transition from an ensemble of random coil states into a more restricted ensemble
of structured states by tuning solution properties and thus the range of electrostatic effects in the structure
with or without the presence of ligand Thus the designed binding interaction was shown to be largely
insensitive to folding unfolding phase equilibria two features essential to SPR signal enhancement Project Narrative
Our research aims to develop a new weapon in the winnable battle for improved food safety an inexpensive
handheld device to optically detect foodborne pathogens within a matter of minutes We combine recent
developments in the disciplines of protein engineering and surface plasmon resonance SPR with off the shelf
electronic components to create a device ideal for proactive monitoring on site eg at a farm food processing
facility or restaurant Although our initial efforts will focus on the detection of Listeria Monocytogenes a
difficult to detect pathogen which is responsible for an estimated deaths annually in the US the principles
underlying our technology can be applied more broadly to the quantitative assay of a broad range of small
molecules proteins viruses and bacteria relevant to public health and food safety

* Information listed above is at the time of submission. *

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