UNCLASSIFIED
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UNCLASSIFIED
Peptide-based Receptor Development for Potential
Integration into Wearable Bio-sensors
Sanchao Liu, Justin Bickford and Matthew Coppock
Sensors and Electron Devices Directorate, CCDC Army Research Lab, Adelphi, MD 20783
Conclusions
Objectives
This work is funded by the Institute for Collaborative Biotechnologies (ICB) through a grant
from the U.S. Army Research Office (ARO), and in collaboration with the Defense Threat
Reduction Agency (DTRA) and the Edgewood Chemical Biological Center (ECBC).
N
3
Target protein
red = epitope of interest
+
Synthetic epitope is
mixed with library of
2 million peptides on
beads
Synthesize epitope
Binding peptides are
identified and then
tested for binding to
full-length target
protein
N
3
N
3
N
3
Peptides that bind to the
epitope in just the right
orientation are covalently
coupled via a click
reaction
N
3
N
3
N
3
Detection
label
CLICK
PCC
“Clickable”
groups
No Cu(I)
catalyst used;
epitope acts
as catalyst
Epitope-targeted PCCs
Development Procedures
Extreme Thermo, Bio, and
Chemical Stability
Multifunctional
Embodiments for Advanced
Applications
1/40
th
the Size of IgG
Monoclonal Antibodies
Capable of Paired Reagent
Discovery
Straightforward Incorporation
of Non-natural Amino Acids
High Affinities and Selectivity
Full Antigen and Epitope
Targeting
Easily Functionalized for
Surface Attachment, Tagging
Limited Batch-to-Batch
Performance Variability
On Demand Scale-up
Through Robotic Methods
PCC
Agents
Targets: Toxins, biothreats,
performance and health biomarkers
IL-17: an inflammation-associated interleukin
IL-6: inflammation and infection responses
IL-10: an immunosuppressive cytokine
IL-11: a signaling molecule improving platelet recovery after chemotherapy
UCHL-1: a human biomarker signaling traumatic brain injury (TBI)
CHIKV E2: virus for high fever
TNFα: Tumour Necrosis Factor alpha
IFN: Interferon gamma, inhibit viral replication
In the multifaceted operational environments of future wars, Army forces will
need to be more adaptive and optimized for execution. The development and
production of ruggedized biosensors can provide real-time feedback for soldier
health, performance, and biothreats.
Soldier wearables
Coppock et al. Sens. Biosensing Res., 2019, 22, 100248.
Coppock et al. Methods, 2019, 158, 12-16.
Coppock et al. Peptide Science, 2017, 108: e22934.
Lai et al., Chem. Eur. J. 2018, 24, 3760.
Agnew et al. Chem. Rev., 2019,
https://doi.org/10.1021/acs.chemrev.8b00660
Advantages of Protein Catalyzed
Capture Agents (PCCs) based Receptors
Cyclic PCC Stability
References
Technology to support:
• Water and food defense
• Individual soldier protection
• Collective protection
• Soldier health monitoring
Meeting requirements for:
• Robust functionality under
environmental extremes
• Suitability for new and
emerging threats
• Capability for rapid
development and production
WILL
PROVIDE
Molecular Recognition is the Heart of the Biosensor
http://dx.doi.org/10.5772/52330
Thermal Stability
Surface Modification on Silicon
Wafer
Biological Stability
@90
o
C
for 1h in
PBS
ADIBO BCN
Biotin Azide
Strep Peptide Azide
PCC Azide
Biotin
Amine
Carboxylic
Acid
• Peptide-based receptors fulfill the need for
alternative antibodies by addressing critical gaps
in adaptability, manufacturability, and stability.
• PCCs can be integrated into multiple platforms for
real-time monitoring of biothreats and soldier
health/performance
• Universal, wearable, disposable, and low cost
Streptavidin coated fluorescent beads
Developing Biosensors for the Future Warfighter
Streptavidin coated magnet beads
What’s next: silicon surface will be
modified with different silanes together
with photolithography procedures to
form different patterns. PCC-based
receptors will then be attached on the
Si-wafer surface.
IL-17F biligands Bi-L
F
Cyclic PCCs showed much higher thermal and biological
stability compared to antibodies
HPLC data for Bi-L
F
before and
after treatment with trypsin for 1h
at 37
o
C
* - Degradation products
Luminex assay of the binding affinity
of PCCs before and after heating at
90
o
C for 1 h.
Rb Anti-E2: antibody
CHIKV E2 ligand: WIYYI
L-amino acids
D-amino acids
Multiple epitopes can be targeted and the resulting PCCs can be
tethered to exploit cooperativity. The bi/tri-ligands show strong
affinity (pM) and high selectivity for the target.
Applications: PCCs will be integrated into silicon-based PIC (photonic
integrated circuits) sensors to provide multiplex detection with true
portability and high accuracy for DoD relevant applications.
