Rapid Detection of Viruses - The Use of Accelerated Molecularly Imprinted Polymers (AMIPs) for the Rapid Detection of Emerging Viral Outbreaks ...
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Rapid
Detection
of Viruses
The Use of Accelerated
Molecularly Imprinted
Polymers (AMIPs) for
the Rapid Detection of
Emerging Viral Outbreaks
April 1, 2020
CSE:SIXW | OTC: ATURF | FSE:AHUH
sixthwave.com
Disclaimer
The data furnished in connection with this proposal is deemed by Sixth Wave Table of Contents
Innovations Inc. to contain trade secrets and commercial or financial infor-
mation, which is privileged and confidential under Title 5, United States Code,
Section 552. Accordingly, such data shall not be disclosed, duplicated, used, or 04 Background
disclosed in whole or in part for any purpose other than to evaluate this pro- 04 The Threat of Viral Outbreaks
posal. 04 SARS-CoV-2
05 SARS-CoV-2 Regions of Interest
07 Techniques for Rapid Detection
07 Recognition at the Molecular Level, Technology Portfolio
10 Proposed Work
10 General Protocol for Platform for Development of RDT for New Threat
11 Principal Components of the RDT System
11 Target Analytes
13 Device Design
16 Signaling, Detection, and Molecular Tags
17 Development Timeline
18 Target Performance
18 Proposed Workflow Progression and Project Milestones
20 References
22 Key Personnel Resumes
22 ARISTOTLE G. KALIVRETENOS
24 JONATHAN P. GLUCKMAN
26 GUNEET KUMAR
28 GLEN E. SOUTHARD
30 DAE JUNG KIM
31 LOUIS W. REICHEL
33 GARRETT M. KRAFT
34 BRANDI C. MAULL
Sixth Wave Innovations Inc.
Rapid Detection System
Background
The SARS-CoV-2, a betacoronavirus, forms a clade within the subgenus sarbecovirus of the
Orthocoronavirinae subfamily. 2 The severe acute respiratory syndrome coronavirus (SARS-
CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) are also betacoronavi-
ruses that are zoonotic in origin and have been linked to potential fatal illness during the
The Threat of Viral Outbreaks outbreaks in 2003 and 2012, respectively.3,4 Based on current evidence, pathogenicity for
SARS-CoV-2 is about 4%, which is significantly lower than SARS-CoV (10%) and MERS-CoV
Viral pandemics are an emerging threat to national security and the global economy. Ex-
(40%). However, SARS-CoV-2 has potentially higher transmissibility (R0: 1.4–5.5) than both
amples of recent viral outbreaks include coronavirus (COVID 2019, MERS 2012, and SARS
SARS-CoV (R0: 2–5) and MERS-CoV (R0:
Sixth Wave Innovations Inc.
Rapid Detection System
The overall structure of SARS-CoV-2 S resembles that of SARS-CoV S, with a root mean Techniques for Rapid Detection
square deviation of 3.8 Å over 959 Cα atoms.7 Similarly SARS-CoV-2 S shares 98% sequence
identity with the S protein from the bat coronavirus RaTG13.9 Even with it’s structural sim- A rapidly developing field for quickly testing patients for infection is Rapid Diagnostic
ilarities to SARS-CoV S, the SARS-CoV-2 S has a 10-20 fold higher binding affinity to ACE2 Tests (RDTs). RDTs are extremely useful as a screening tool to process large numbers of pa-
at ~15nM.7,10 The overall structural similarity between the SARS-CoV and SARS-CoV-2 spike tients, often designed to be administered by unskilled technicians, these tests are fast, ac-
protein has led to testing of SARS-CoV RBD-directed monoclonal antibodies (mAbs) for curate, and inexpensive relative to other techniques. As a first line of screening these tests
cross-reactivity to the SARS-CoV-2 RBD. Cross-reactivity of the SARS-CoV RBD-directed can be over 90% accurate and quickly determines if the patient warrants the expenditure
mAbs has shown almost no affinity to SARS-CoV-2 RBD despite the relatively high degree of more resources. If positive, the result is validated by a second technique and the nec-
of structural homology.7 Having a flexible platform for diagnostics that can easily be ad- essary medical resources are deployed. RDTs are generally developed from immunoassay
justed for small structural changes and genetic drift is essential to give healthcare work- techniques and have been used in pregnancy tests, drug tests, HIV detection, Influenza
ers tools in real time to effectively mitigate outbreaks. detection, strep throat diagnosis, and many others. Immunoassays use antibody/antigen
complexes as their primary mechanism of detection. The antibodies can be developed to
complex with proteins, DNA, or RNA as is widely performed in the clinical tests of Western
blots, Southern blots, and Norther blots, respectively. Similarly, there is a technique of
using DNA or RNA as the complexing agent known as Aptamers.11–14 This technology uses a
specific DNA or RNA sequence, which is determined by screening a library of nucleic acid
sequences against the target analyte, to selectively bind to the target. Another form of
virus detection is Polymerase chain reaction (PCR). This technique is more time and cost
intensive because it requires isolation and sequencing of the virus’ genome to make an
accurate diagnosis. This is a common second technique used to validate a positive test
from ammitial RDTs.
Recognition at the Molecular Level, Technology Portfolio
Sixth Wave specializes in the detection and binding of target analytes at the molecular
level. Sixth Wave has successfully developed and commercialized technology for the de-
tection of explosives, biogenic amines, and selective binding and adsorption of target
plant metabolites and metal complexes. Sixth Wave has a technology portfolio consisting
of several different mechanisms for detecting target molecules at the nanometer level.
The flagship technology, which will be a part of the technology review for this application,
uses a branch of nanotechnology called “MIPs” – Molecularly Imprinted Polymers. The se-
lective binding of MIPs is well known in the literature and have even been referred to as
“plastic antibodies”. The MIPs can be engineered into many forms and can be suspended
Figure 2: Structure of SARS-CoV-2 S in the prefusion conformation in an assay, adhered to a wipe, or fixed to a membrane for lateral flow tests. These materi-
(A) Schematic of 2019-nCoV S primary structure colored by domain. Domains that were excluded from the als can be specially designed to form a flexible platform that can continuously be altered
ectodomain expression construct or could not be visualized in the final map are colored white. SS, signal
to meet the demand of detecting constantly evolving viral strains.
sequence; S2′, S2′ protease cleavage site; FP, fusion peptide; HR1, heptad repeat 1; CH, central helix; CD,
connector domain; HR2, heptad repeat 2; TM, transmembrane domain; CT, cytoplasmic tail. Arrows denote
protease cleavage sites. (B) Side and top views of the prefusion structure of the 2019-nCoV S protein with
a single RBD in the up conformation. The two RBD down protomers are shown as cryo-EM density in either
white or gray and the RBD up protomer is shown in ribbons colored corresponding to the schematic in (A).
06 07
Sixth Wave Innovations Inc. Rapid Detection System “MIPs” – Molecularly Imprinted Polymers MIPs are polymers designed to be highly selective for a specific analyte. MIPs are prepared by polymerizing a ligand(s) which coordinates or non-covalently “binds” with the imprint. The target molecule and the polymerizable ligand are incorporated into a pre-polymer- ization mixture, allowed to interact and pre-organize, then polymerized (typically in the presence of one or more non-crosslinking monomers and a cross-linking monomer) to form a porous substrate with 3-dimensional cavities and chemistries complementary to the complexed imprint. The target analyte thus acts as a “template” to define a cavity or binding site within the polymerized matrix which is specific to the target analyte (e.g., has a shape or size corresponding to the target molecule). The desired target analyte itself may be utilized to form the MIP or a “surrogate” may be used which mimics the known target. Post-polymerization, the template is then removed from the MIP prior to its use as a selective absorbent for the analyte target of interest. Figure 3: Illustration of the basic components of Molecularly Imprinted Polymers. 08 09
Sixth Wave Innovations Inc.
Rapid Detection System
Proposed Work
Principal Components of the RDT System
The flexible platform would work by having a target analyte (e.g virus), identified by the
medical community, and screening the viral MIP library for the most selective binding
interaction in collaboration with a Virology Laboratory. Once a suitable MIP is selected,
General Protocol for Platform for Development of RDT a compatible and sensitive visualization technique will be selected. In the initial design,
for New Threat a standard protein labeling agent may be employed for visualization of the bound virus.
Once performance is validated (sensitivity/selectivity), the chemistry will be scaled for
fast deployment of first generation RDTs (qualitative testing).
The goal of this proposal is to build a platform device/technology that can be rapidly
applied to new threats once information is available from the medical community. The To improve specificity, the visualization chemistry may be further modified as new infor-
device can be envisioned as a cartridge-type system where the physical format, detection mation becomes available from the medical community concerning ligands (e.g protein,
method and sampling/detection process is the same for all threats. The interchangeable antibody, small molecule) that show specific affinity for the virus. Once several useful
piece would be the molecular recognition chemistry which would be unique to each new candidates are identified for the target, an array of the ligands modified with molecular
threat. For a viral threat such as SARS-COV-2, recognition of the intact virus15 would be the tags can quickly be synthesized and screened for performance (selectivity and sensitivity).
most efficient process timewise. This may be done via the availability of a MIP that recog- Once validated, the process will be scaled for fast deployment of second generation RDTs
nizes/binds the virus. Clearly there is not time to develop a novel MIP after a new outbreak with enhanced specificity.
has occurred. To avoid these delays and rapidly develop a new diagnostic, we propose to
build a family of MIPs based on the general shape, size and morphology of generic class- One of the major advantages of our technology is that it can utilize off the shelf known
es of viruses. These would be prepared utilizing virus surrogates and/or attenuated or chemistries and synthetic ligands for the labeling and visualization of the MIP-bound
inactivated viruses (non-infectious). The MIPS would be part of an inventory ready to im- virus that don’t require animal hosts for production. This further accelerates the develop-
mediately be deployed for screening against a new threat. The most selective MIP would ment timelines. Furthermore, the use of known chemistries and synthetic molecules will
be further incorporated into a previously defined format for use in the diagnostic plat- enhance stability and reduce failed tests due to degradation of biomolecules like anti-
form system. As the MIP chemistry is already known, direct scale-up to manufacture could bodies. The use of synthetic ligands also allows for simple modifications in cases where
be initiated immediately and applied to kits that are already able to be mass produced. the virus mutates. With antibody-based systems, the labeled antibody will lose its utility
if the virus mutates and the binding affinity is lost.
Potential Rapid Colorimetric Test Format
Target Analytes
The primary target for this technology would be the entire virus particle. However, this
technology also has the capacity to target other biomarkers15 (such as DNA, RNA, protein,
antibodies, or small molecule) to optimize selectivity and performance. Many infection
diagnostic tests rely on the detection of antibodies produced as a result of the infection.
This is an example of indirect recognition since it isn’t detecting the virus directly but
rather the body’s response (the antibody) to the viral infection. Direct recognition would
be the detection of the virus or a viral component such as its DNA or RNA sequence or a
specific protein. Direct recognition allows for earlier detection since the virus is being de-
Figure 4: Schematic of a SARS-CoV-2 detection test using Sixth Wave MIP technology. tected directly. In the case of indirect recognition and antibody detection, it can take up to
30 days after infection for the body to ramp up antibody levels to the point of detectable
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Rapid Detection System
limits. However, direct recognition has reduced efficacy late in the infection when recovery
is underway. Under these conditions the body has mounted a sufficient immune response
reducing the concentration of virus particles in the body. Here indirect recognition would
be preferred because the elevated levels of antibodies will remain in the system well after
the infection has been eliminated. In general, Sixth Wave would strive to develop an RDT
test using direct recognition mechanisms for the viral target to prioritize diagnosing new
infections to help contain the spread of the viral agent. After release of an initial RDT for
the virus, development would continue to add detection capabilities for the antibody to
the diagnostic test as well using a more traditional immunoassay protocol to detect the
presence of IgM and IgG antibodies identified for the novel virus. This would allow medical
professionals to have two to three independent markers for determining infected patients
and give bracketed ranges of where they may be in the infection processes (early – no an-
tibodies detected, or late – no virus detected).
Device Design
The principle of virus detection based on antigen/antibody recognition is divided into
three stages: 1) immobilization of antibody on a substrate, 2) migration of viruses to an-
tibody, and 3) binding of viruses and antibody. Sixth Wave MIP technology has the same
recognition capabilities and can directly replace the antibody in this binding interaction.
In the present proposed diagnostic, the virus is bound to the Sixth Wave MIP via a highly
selective molecular recognition process. Virus detection is achieved by various sensing
techniques based on the binding event. Three suggested detection approaches compat-
ible with MIPs include: 1) quartz crystal microbalance (QCM) sensor, 2) lateral flow immu-
noassay, and 3) electrochemical sensor.
A QCM sensor is a kind of mass sensitive sensor. The basic material of the QCM sensor con-
sists of quartz crystal, which is equipped with metal electrodes (e.g. gold). A sensor surface
is modified with coating of MIPs, which is used to enable detection of viruses. An appro-
priate electronic circuit is necessary to make conversion of the measured virus quantity to
an electrical signal. The basic working principles of the quartz crystal microbalance sensor
are displayed in Figure 5. Viruses that are present in the surrounding solution of a QCM
sensor will interact with the sensor surface. In this interaction, viruses are bound to the
MIP. The bound virus results in a mass change on the sensor surface. Consequently, the
mass change on the sensor surface is converted to a frequency change. A QCM sensor is a
useful tool for biomolecule detection and it has advantages such as high sensitivity, low
response time, continuous operation, portable device, label-free, and real-time detection
ability. QCM biosensor systems are composed of three components which are receptor,
transducer, and signal monitoring system.
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Rapid Detection System
color of the reaction pad is changed by adsorption of virus onto the MIPs and activation
of a correlating molecular tag.
Electrochemical biosensors have electrodes which translate the chemical signal into an
electrical signal. Electrochemical sensor can detect various biomolecules in the human
body such as glucose, cholesterol, uric acid, lactate, DNA, hemoglobin, blood ketones, and
viruses. Electrochemical biosensor typically requires a working electrode, a counter (or
auxiliary) electrode and a reference electrode. The reference electrode is maintained at
a distance from the site of the biological recognition element and analyte interaction to
establish a known and stable potential. The working electrode acts as the transduction
component when the interaction occurs whereas the counter electrode measures current
and facilitates delivery of electrolytic solution to allow current transfer to the working
electrode. The working and counter electrodes should be conductive and chemically sta-
ble and are mostly composed of carbon or inert metals like gold and platinum whilst the
reference electrode is typically silver or silver chloride. A schematic diagram of an electro-
chemical biosensor is presented in Figure 7.
Figure 5: Basic working principle of a quartz crystal microbalance (QCM) sensor.
The lateral flow immunoassay is a colori-
metric platform, which is normally used
in determining a biomolecule or a chem-
ical element in a solution with the aid of
a color reagent. This method is widely ap-
plied to the analysis of solution samples
in medical or industrial fields. It enables
the detection of target molecules by the
naked eye without any sophisticated in-
struments. A lateral flow dipstick is pri-
marily used with immunoassays. Figure 6
shows the configuration of a typical lat-
eral flow dipstick. The lateral flow dip-
stick (LFD) is composed of a sample pad,
conjugate pad, reaction pad, wicking pad,
and plastic card. For virus detection, MIPs
Figure 7 A schematic diagram of an electrochemical biosensor. To enhance the detection function, the sur-
are coated on the conjugation pad and face of the working electrode is modified with MIPs.
the reaction pad. The biotinylated bovine
Figure 6: Configuration of a lateral flow dipstick
serum albumin is doped onto the nitro-
cellulose membrane of the reaction pad.
During the detection process, the surface
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Signaling, Detection, and Molecular Tags The labeling process would be similar to antibodies, with the use of colloidal gold or dye
conjugates to add a detectable group.
The choice of pathway for the signaling/detection of a selective binding event of the
viral target to the MIP will be dependent on the device design. Process 3: The presence of the virus may be determined directly by the binding of the virus
to a labeled synthetic ligand (nature of ligand determined by medical/research commu-
QCM: No additional modification is required as detection is mass sensitive. nity). If it is a small molecule or peptide, the use of a dye conjugate would be the most
advantageous to add a detectable group.
Electrochemical Biosensor: No additional modification is required as detection is de-
pendent on electrical change Process 4: If no known ligands or other biological entities are known or available during
initial RDT development, the presence of the virus may be visualized utilizing known
Colorimetric/Fluorescent Devices: amine/protein labeling techniques. Known chemistries are highly reactive and provide a
high level of sensitivity based on the dye choice although they are not specific for the tar-
For lateral flow and microtiter plate type devices, the presence of the analyte to be detect- get virus. The highly selective nature of the MIP/virus binding will minimize false positive
ed must trigger a binding event which induces an observable colorimetric or fluorescent results using this process.
change. In the Sixth Wave viral RDT, the target virus analyte is bound selectively to the MIP
during sample addition and incubation. In order to achieve a color change, the bound Indirect Detection of the Virus:
analyte will form a conjugate with an entity which contains a tag. The nature of this tag is
dependent on the entity used for binding. The presence of the virus may be determined indirectly by the binding of an anti-virus
antibody (IgG and IgM as per the medical community) to a labeled antigen entity (deter-
Direct Detection of Virus: mined by medical/research community). The labeling process would be similar to the
above methods, depending on the nature of the antigen entity.
Process 1: The presence of the virus may be determined directly by the binding of the vi-
rus to a labeled antibody conjugate. There are two readily available options to prepare a Development Timeline
labeled antibody.
The key benefit to Sixth Wave’s technology is the potential to finish development and
In one embodiment, the antibody is adsorbed on gold nanoparticles (colloidal gold) which scale quickly. This is due to the fact that immunoassay based RDTs are reliant on animal
provides a distinct red to orange color change depending on the gold particles utilized. surrogates for the production of new antibodies which may take 2-3 months. Initial design
This is a well-known process and is common in lateral flow and other immunoassays. A and development of the flexible platform will take 6-12 months, but after the platform has
useful feature is it is applicable to any protein, enzyme antibody. been developed, new product development would follow the table listed below.
Alternatively, the antibody may be conjugated to a chromophore or fluorescent entity
Steps for RDT Development Sixth Wave Immunoassay
using standard protein conjugation chemistry (e.g. water soluble carbodiimide couplings,
conjugation of activate ester forms of chromophores/fluorophores). A potential extension New Virus Identification and Characterization 1 Month 1 Month
of this chemistry is to utilize an amine terminated dendrimer as the detection group con- Development of new RDT 1-2 Months 3-6 Months
jugated to the antibody. In this format, the dendrimer can be pre-labeled with multiple
Clinical Validation of RDT 1 Month 1 Month
dye molecules which may enhance sensitivity.
Process 2: The presence of the virus may be determined directly by the binding of the
virus to a labeled virus binding protein (determined by medical/research community).
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Target Performance and will be dependent on the nature of the polymer matrix synthesized. This process will
require intensive work initially, with large number of potential MIPs being screened, but
Detection limit: 1 ng/mL the work will diminish as the MIP synthesis progresses from new MIP synthesis to formula-
Time: less than 30 minutes tion optimization. During the formulation optimization there will be no change in labeling
Detection: Colorimetric and binding chemistry but may require a few adjustments to achieve the correct level
Specificity and Sensitivity: 90% of detection with new formulations. This process is expected to be time intensive for six
weeks then requiring diminishing input over the next 2-4 weeks, potentially requiring a
Proposed Workflow Progression and Project Milestones final optimization at the very end of MIP synthesis.
Initial work on the virus detection platform would be focused on using SARS-COV-2 as a After the MIP has been synthesized the next stage is to process it to be compatible with
case study for design principles and a broader scope to be able to detect different virus the target device or platform. This target will be in mind the entire time during MIP syn-
classes would be pursued after achieving a working prototype for SARS-COV-2. The project thesis and will allow for quick transition from MIP synthesis to prototyping. The key deliv-
would start by sourcing the attenuated virus and spike protein from a supplier. The virus erable is to produce a device that delivers within the specifications determined, and that
and spike protein represent the two best imprint candidates. it can produce reproducible results. Furthermore, the MIP formulation can be applied to
multiple devices and give reproducible results.
Lab work would begin with the development of MIPs for the two imprint candidates. For
each candidate, a matrix will be developed containing monomers which have function- The final phase is testing of clinical samples. Here samples of patient’s body fluids will
ality complimentary to the candidate to provide an active binding pocket in the final be tested with the prototype devices and tested for accuracy, sensitivity, selectivity, and
MIP, not just dependent on shape and size of the imprint candidate. These may include speed. Training requirements for technicians will also be determined at this stage, but
charged monomers, uncharged polar monomers and uncharged hydrophobic monomers. with a colorimetric design there should be minimal training time required. Successful
As per Figure 3 above, a pre-polymerization mixture consisting of the active monomers, clinical validation will lead to scale-up and mass production with an appropriate partner
the backbone core monomers, crosslinking monomer and the imprint candidate will be that was involved in the prototyping stage as well.
incubated to pre-organize followed by polymerization to form the MIP. During subsequent
processing the imprinted material will be removed, and the empty MIP utilized for testing.
A series of polymers will be prepared with the goal of providing the strongest and most
selective binding pocket for the imprint candidate.
Once synthesized and processed the MIPs will be tested in non-competitive and compet-
itive binding experiments to determine capacity, robustness and selectivity. These exper-
iments will give a general idea about the performance parameters of the MIPs and give
a feedback loop for optimizing the MIP formulation. The new synthesis and optimization
feedback loop are estimated to take at least 4 months to produce a MIP that will meet the
criteria listed in the target performance section.
In parallel with the synthesis of new MIPs will be the development of the labeling and
binding chemistry. Once an imprint has been selected and there is a set of ligands show-
ing preliminary affinity towards the imprint, the specific chemistry for the molecular tags
can be determined. This chemistry is dependent on the nature of the imprint. Similarly,
the binding of the recognition site, the MIP, to the device substrate will also be required
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Ph.D. Organic Chemistry, Colorado State University (1990) Modulation” Agarwal, P.K.; Schultz, C.; Kalivretenos, A.; Ghosh, B.; Broedel,
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WORK EXPERIENCE PATENTS
Sixth Wave Innovations Inc., January 2020 to present, Director of Science 1) “Butyryl-Tyrosinyl Spermine, Analogs thereof and Methods of Preparing and
Aurora Analytics, LLC, 2004 to present, COO & Using Same” Koji Nakanishi, Danwen Huang, Soek-Ki Choi, Aristotle Kalivretenos
Managing Member and Robert Goodnow, US Patent # 6,001,824, 1999.
Butler Manufacturing LLC, 2014 to present, 2) “Detection of Analytes by Fluorescent Lanthanide Metal Chelate Complexes
Consultant Containing Substituted Ligands” Arthur E. Colvin, George Y. Daniloff,
Notre Dame of Maryland University, 2005 to 2008, Adjunct Professor of Chemistry Aristotle G. Kalivretenos, David Parker, Edwin E. Ullman and Alexandre V.
Senseonics, Inc., 1999 to 2004, Scientist II. 1998-1999, Scientist Nikolaitchik, US Patent # 6,344,360, 2002.
Paragon Bioservices, Inc., 1998, Consultant 3) “Detection of glucose in solutions also containing an alpha-hydroxy acid or a
Univ of Maryland Baltimore County, 1998 to present, Adjunct Professor of Chemistry. 1992 beta-diketone” George Y. Daniloff, Aristotle G. Kalivretenos, and Alexandre V.
to 1998, Assistant Professor of Chemistry Nikolaitchik, US Patent # 7,078,554, 2006.
Unilever Research, U.S., 1991-1992, Synthetic Consultant 4) “Amine detection method and materials” Aristotle G. Kalivretenos,
Columbia University, 1990 to 1992, Post-doctoral Research Fellow US Patent # 7,229,835, 2007.
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SELECTED PUBLICATIONS US Patent # 7,592,183, 2009.
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L. S. J. Org. Chem. 1991, 56, 2883-2894. American Chemical Society (1986)
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3) “Labeling Studies of photolabile philanthotoxins with nicotinic acetylcholine
receptors: mode of interaction between toxin and receptor” Choi, S.-K.;
Kalivretenos, A.G.; Usherwood, P.N.R.; Nakanishi, K. Chemistry & Biology 1995, 2, 23.
022 023Sixth Wave Innovations Inc.
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JONATHAN P. GLUCKMAN, Ph.D. 5) Gluckman, J.G. & Becker, D. (1993). “Knowledgeable Observation Analysis-Linked
Advisory System (KOALAS) and Multi-sensor Integration.” (Technical Report
No. N1808-TR-93-00151) Naval Air Warfare Center (1993).
EDUCATION
Ph.D. Human Factors/Experimental Psychology, University of Cincinnati (1990)
HONORS
M.A. Human Factors/Experimental Psychology, University of Cincinnati (1988)
Outstanding Service to the Shuttle Cockpit Upgrade Team, 1998, Air Vehicle and Crew
B.S. Psychology, Bradley University (1985)
Systems Department Scientific Award, 1993.
WORK EXPERIENCE
PATENTS
Sixth Wave Innovations Inc., July 2013 to present. Chairman & CEO
Southard, G. E.; Gluckman, J. P. Molecularly Imprinted Polymer Beads for Extraction of
Raptor Detection Technologies, LLC, July 12, 2010 to June 26, 2013. General Manager & EVP
Metals and Uses Thereof. U. S. Patent 9,504,988, 2016.
Raptor Detection, Inc. April 2007 to 2010. President & CEO
Link Plus Corporation. December 2004 to April 2007. President & Chief Operating Officer.
Macklin, J.D., Bonneau Jr., W.C., Gluckman, J.P., Dorovskoy, I, Security Polymer Threat De-
January 2004 to December 2004. Executive Vice- President & Chief Operating Officer.
tection Distribution System, 2013.
Integrated Dynamics Inc., 1996 to 2003. Chief Executive Officer & Co-founder.
JS Technologies, Corp. a subsidiary of Integrated Dynamics, Inc., 2001 to 2003.
Vice President Technology/Chief Technical Officer & Co-founder.
PROFESSIONAL SOCIETY MEMBERSHIPS
JJM Systems, Inc., Intelligent Control Technologies Division, 1993 to 1996. Division
American Chemical Society (2014)
Manager.
Society of Mineralogical Engineering (2014)
US Naval Air Warfare Center, Aircraft Division. 1991 to 1994. Block Manager / Staff
Engineering Psychologist.
MAJOR PUBLICATIONS
1) Molecularly Imprinted Polymer Applications for the Gold Mining Industry,
G. Southard, J. Gluckman, B. Maull Proceedings of Heap Leach Solutions, 2015
September 14-16, 2015, Reno, Nevada, USA.
2) Gluckman, J.G., Warm, J.S., Dember, W.N., & Rosa, R.R. “Demand Transitions
and Sustained Attention”. Journal of General Psychology. (1993)
3) Harris, S.D., Ballard, L., Girard, R. Gluckman, J.P.“Sensor Fusion and Situation
Assessment: Future F/A-18 Capabilities.”In A. Levis and I. Levis (Eds.), Science of
Command and Control: Part III Coping with Change, AFCEA International
Press,(1994).
4) Harris, S.D. and Gluckman, J.P. (1994) “A Conceptual Framework for Advanced
Tactical Information Management Systems.” In A. Levis and I. Levis (Eds.), Science
of Command and Control: Part III Coping with Change.
024 025Sixth Wave Innovations Inc.
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GUNEET KUMAR, Ph.D. 4) “Enzymatic grafting of a natural product onto chitosan to confer water solubility
under basic conditions”, G. Kumar, P.J. Smith, and G.F. Payne, Biotechnology and
Bioengineering, Vol 63, No. 2, 154-165 (1999).
EDUCATION
5) “Plant cell biodegradation of a xenobiotic nitrate ester, nitroglycerin”, A. Goel,
G. Kumar, G.F. Payne, and S.K. Dube, Nature Biotechnology, Vol 15, 174-177(1997).
Ph.D. Chemical Engineering, March 1994
6) “Polyoxazoline-Peptide Adducts that Retain Antibody Avidity”, W.H. Velander,
Virginia Polytechnic Institute & State University, Blacksburg, Virginia, U.S.A.
R.D. Madurawe, A. Subramanian, G. Kumar G. Sinai-Zingde, J.S. Riffle, and C.L.
M.Sc. Organic Chemistry, May 1986, Indian Institute of Technology, Kharagpur, India
Orthner, Biotechnology and Bioengineering, Vol. 39, 1024-1030 (1992).
B.Sc. Chemistry, Spring 1984, Indian Institute of Technology, Kharagpur, India
PATENTS
WORK EXPERIENCE
1) “Production of Bio-Butanol and related products” P. Paripati, G. Kumar, 2014 :
Patent application # 12825219.
Sixth Wave Innovations Inc., Senior Scientist (2020-current)
2) “Corecovery of Bio-Oil and fermentable sugars from oil bearing biomass”,
Aurora Analytics, Senior scientist/process development (2019)
Severa G., Kumar G., Cooney M., Filed provisional Patent 2013,
Suganit Systems Inc. (2006 – 2016) VP, Bio-Renewable Technologies
Application # 61/818,393.
In Vitro Technologies., Inc. (August 2005-July 2007) Sr. Scientist
3) “Biomass Pretreatment” Varanasi, S., C. Schall, A.P. Dadi, J. Anderson, K. Rao,
ChemPacific & UPM Pharmaceuticals, Consultant (2004-2005)
P. Paripati, and G. Kumar. Patent No. 8,546,109.
Bio Scale-up Facility, University of Maryland, College Park (2002-2004) Protein Purifica-
4) “Pretreatment of Biomass” Varanasi, S., C. Schall, A.P. Dadi, J. Anderson, K. Rao,
tion Specialist
P. Paripati, and G. Kumar. Patent No. 8,030,030.
Bio Scale-up Facility and Center for Agricultural Biotechnology, University of Maryland,
5) “Modified Chitosan Polymers and Enzymatic Methods for the Production
College Park. (May 1999 – March 2002) Faculty Research Associate
Thereof”, G.Kumar and G.F. Payne. Patent No. 7,288,532 .
Dept. of Chemical and Biochemical Engineering, University of Maryland at Baltimore
County (UMBC).(June 1994 - June 1995, October 1996 - April 1999) Post-Doctoral Fellow,
Research Associate
PROFESSIONAL SOCIETY MEMBERSHIPS
AICHE AND ACS 1995
SELECTED PUBLICATIONS
1) “Co-recovery of lipids and fermentable sugars from Rhodosporidium toruloides
using ionic liquid co-solvents: Application of recycle to batch fermentation.,”
Godwin Severa, Kumar, G., and Michael J. Cooney. 2014, Biotechnology Progress.
2) “Co-recovery of bio-oil and fermentable sugars from oil-bearing biomass”.
Godwin Severa, Kumar, G., and Michael J. Cooney. International Journal of
Chemical Engineering. Volume 2013 (2013), Article ID 617274,
http://dx.doi.org/10.1155/2013/617274
3) “Enzymatic gelation of the natural polymer chitosan” G. Kumar, J.F. Bristow,
P.J. Smith, and G.F. Payne, Polymer, Vol 41, 2157-2168 (2000).
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GLEN E. SOUTHARD, Ph.D. 2007, 581(2), 202-207.
5) Soluble and Processable Phosphonate Sensing Star Molecularly Imprinted
EDUCATION
Polymers, Southard, G. E.; Van Houten, K. A.: Murray, G. M. Macromolecules 2007,
40, 1395-1400.
Ph. D. Inorganic Chemistry, University of Michigan (1998)
6) Heck Cross-coupling for Synthesizing Metal Complexing Monomers, Southard,
B. S. Chemistry, Iowa State University (1991)
G. E.; Murray, G. M. Synthesis 2006, 15, 2475-2477.
7) Molecularly Imprinted Ion Exchange Resin for Fe3+, Owens, G. S.; Southard,
WORK EXPERIENCE
G. E.; Murray, G. M. Sep. Sci. Tech. 2005, 40, 2205-2211.
8) Synthesis of Vinyl-Substituted β-Diketones for Polymerizable Metal Complexes,
Sixth Wave Innovations Corp., 2014 to 2017. Chief Scientific Officer
Southard, G. E.; Murray, G. M. J. Org. Chem. 2005, 70(22), 9036-9039.
Bard Access Systems, Utah, 2010 to 2014, Consultant
9) Molecularly Imprinted Polymers, George M. Murray and Glen E. Southard,
University of Tennessee Space Institute, 2010 to 14, Consultant
Molecularly Imprinted Materials: Science and Technology, Yan and Ramstrom,
Environmental Protection Agency, 2009 to 2014, Peer Reviewer—reviewed business pro-
eds., Marcel-Dekker, Nov. 30, 2004.
posals for technical & commercial merit for EPA Small Business Innovative Research
10) Development of Molecularly Imprinted Polymer Sensors for Chemical Warfare
(SBIR) program and Science to Achieve Results (STAR) Program, reviewed graduate stu-
Agents, Boyd, J. W.; Cobb, G. P.; Southard, G. E.; Murray, G. M. JHUAPL Technical
dent research submissions for technical merit as criteria for EPA funding,
Digest, 2004, 25(1), 44-49.
MIP Solutions, Inc., 2004–2008, MOB of Directors, Chief Scientific Officer and Laboratory
Manager.
The Johns Hopkins University Applied Physics Laboratory, 2001–2004, Senior Profession-
PATENTS
1) Southard, G. E.; Gluckman, J. P. Molecularly Imprinted Polymer Beads for Extraction
al Staff, Polymer Chemist in Technical Services Department.
of Metals and Uses Thereof. U. S. Patent 9,504,988, 2016.
Schafer Corporation – Schafer Laboratories, 1999 – 2001, Scientist 3.
2) Cox, J. B.; Southard, G. E.; Messerly, S. Ruggedized ultrasound hydrogel insert. U. S.
Eastern Michigan University – Coatings Research Institute, 1998–1999, Post-Doctoral Fel-
Patent 9,211,107, 2015.
low.
3) Southard, G. E.; Van Houten, K.; Murray, G. M. Process for Preparing Molecularly
Imprinted Polymer Ion-Exchange Resins. U. S. Patent 8,085,208; 2011.
MAJOR PUBLICATIONS
4) Southard, G. E.; Murray, G. M. Processable Molecularly Imprinted Polymers.
1) Molecularly Imprinted Polymer Applications for the Gold Mining Industry,
U. S. Patent 7,678,870; 2010.
G. Southard, J. Gluckman, B. Maull Proceedings of Heap Leach Solutions, 2015
5) Southard, G. E. Molecularly imprinted polymers (MIPS) for the selective removal
September 14-16, 2015, Reno, Nevada, USA.
of inorganic contaminants from liquids. U. S. Patent 7,476,316; 2009.
2) Southard, G. E. and Murray, G. M., “Molecularly Imprinted Polymer Receptors for
6) Southard, G. E. Sensor for Monitoring an Analyte. U. S. Patent 7,319,038; 2008.
Sensors and Arrays,” in Principles of Bacterial Detection: Biosensors, Recognition
7) Southard, G. E.; Murray, G. M.; Ko, H. W. Neutron detection based on boron
Receptors and Microsystems, Zourob, M., Elwary, S. and Turner, A. Eds., Springer
activated liquid scintillation. U. S. Patent 7,126,148; 2006.
Verlag, 2010.
8) Southard, G. E.; Murray, G. M. Process for preparing vinyl substituted
3) Synthesis and Spectroscopic Characterization of Molecularly Imprinted Polymer
beta-diketones. U. S. Patent 7,067,702; 2006.
Phosphonate Sensors, G. E. Southard, K. A. Van Houten, Edward W. Ott Jr. and
G. M. Murray, Polymers and Materials for Anti-Terrorism and Homeland Defense,
ACS Symposium Series, Lawson, G. E. and Reynolds, J. G., Eds., American Chemical
PROFESSIONAL SOCIETY MEMBERSHIPS
American Chemical Society (1993)
Society, Washington, D. C., 2008.
Society of Mineralogical Engineering (2014)
4) Luminescent Sensing of Organophosphates Using Imprinted Polymers Prepared by
Utah Technology Council (2007)
RAFT Polymerization, Southard, G. E.; Ott, E. W., Jr.; Murray, G. M. Microchimica Acta
028 029Sixth Wave Innovations Inc.
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DAE JUNG KIM, Ph.D. LOUIS W. REICHEL, Ph.D.
EDUCATION EDUCATION
Ph.D. Chemical Engineering, Ajou University (2002) Ph.D. 1985, Organic/Analytical Chemistry. University of Southern Mississippi.
M.S. Chemical Engineering, Chonnam National University (1995) M.S. 1983, Organic Chemistry. University of New Orleans.
B.S. Chemical Engineering, Chonnam National University (1992) B.S. 1977, Chemistry. University of Maryland.
WORK EXPERIENCE
WORK EXPERIENCE
Sixth Wave Innovations Inc., August 2015 to present. Sr. Scientist
Red Leaf Resources, March 2014 – May 2015. Chemical Engineer Sixth Wave Innovations Inc., Senior Scientist (2020-current)
Utah State University Bioenergy Center, October 2013 – March 2014. Lab Manager Aurora Analytics, LLC, Feb 2009 – 2020. Scientist
UK Center for Applied Energy Research, November 2011 – August 2013. Senior Research Amulet Pharmaceuticals Inc., Jan 2007-May 2008. Research Chemist
Engineer Novavax, Inc.,
University of Utah, November 2007 – October 2011. Research Associate Jan 2006 – July 2006. Senior Scientist, Pharmaceutical Development, Malvern, PA.
Texas Tech University, March 2006 – October 2007. Postdoctoral Fellow Mar 2005 – Jan 2006. Product Development and Formulation Scientist, Rockville, MD and
University of Utah, March 2004 – March 2006. Postdoctoral Fellow Philadelphia. PA.
COCAT, March 2002 – March 2004. Principal Chemical Engineer Mar 2001 – Mar 2005. Director of Quality Control and Analytical Laboratory, Rockville, MD.
Neophotech, February 2001 – February 2002. Principal Chemical Engineer Nov 1998 – Mar 2001. Senior Analytical Chemist, Columbia and Rockville, MD.
General Motors Korea, December 1994 – January 2001. Senior Chemical Engineer Guilford Pharmaceuticals, Baltimore, MD. July 1997 – Nov 1997. Chemist, Product Develop-
Chonnam National University, February 1993 – November 1994. Research Associate ment (contract).
University of Southern Mississippi, Hattiesburg Sep 1993 – July 1997. Postdoctoral Fellow,
PUBLICATIONS AND PRESENTATIONS Polymer Science Department.
Peer-reviewed journal papers: 34, Book chapters: 6, Conference and invited presenta- Zeneca, Inc., Feb 1989 – Aug 1993. Senior Chemist, Agricultural Products Division.
tions: 65, Patents: 3 Geo-Centers, Inc., Nov 1987 – Sep 1988. Organic Synthesis Research Chemist.
Representative Journal Papers University of New Orleans, New Orleans, LA. June 1985 – Nov 1987. Postdoctoral Research,
S. Ritz, J. Gluckman, G. Southard, B. Maull, and D.J. Kim, “Imprinted Resin—The 21st Centu- Department of Chemistry.
ry Adsorbent”, Extraction, 1943-1960 (2018).
D.J. Kim, J. Wang, and M. Crocker, “Adsorption and desorption of propene on a commer- PROFESSIONAL SOCIETY MEMBERSHIPS
cial Cu-SSZ-13 SCR catalyst”, Catalysis Today, 231:83-89 (2014).
D.J. Kim, B. Weeks, R. Pitchimani, D.E. Snow and L.J. Hope-Weeks, “A Simple Method for American Chemical Society
the Removal of Thiols on Gold Surfaces Using an NH4OH–H2O2–H2O Solution”, Scanning,
30:118–122 (2008).
D.J. Kim, B. Weeks and L.J. Hope-Weeks, “Effect of Surface Conjugation Chemistry on the
Sensitivity of Microcantilever Sensors”, Scanning, 29:245-248 (2007).
D.E. Snow, B. Weeks, D.J. Kim, A. Loui, B. Hart and L.J. Hope-Weeks, “Static Deflection Mea-
surements of Cantilever Arrays Reveal Polymer Film Expansion and Contraction”, J. Col-
loid and Interface Sci., 316:687-693 (2007).
030 031Sixth Wave Innovations Inc.
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GARRETT M. KRAFT, Ph.D. 2016.
Kraft, G. M. Biomimetic materials: brushes, foams, and fibers. University of Connecticut
GAANN Fellowship Research Seminars, U.S. Department of Education, Storrs, CT, October
EDUCATION
17, 2016.
Kraft, G. M.; Bento, J. L.; Madugula, D.; Marinez, A.; Adamson, D. H. Synthesis of complex
Ph.D. Polymer Science, University of Connecticut
amphiphilic polymers by azeotropic distillation techniques. Proceedings of the 250th
B.S. Chemistry, Biology, University of Wisconsin-Stevens Point
American Chemical Society National Meeting, Boston, MA, August 16-20, 2015.
Kraft, G. M.; Weinstein, S. D.; Woltornist, S. J.; Adamson, D. H. Infusion of catalytically ac-
WORK EXPERIENCE
tive polymers for templated condensation of metal oxides in foam composites. Proceed-
ings of the 250th American Chemical Society National Meeting, Boston, MA, August 16-20,
Sixth Wave Innovations Inc., July 2017 to present. Senior Scientist
2015.
University of Connecticut, August 2012 to July 2017, Graduate Researcher
Kraft, G. M.; Woltornist, S. J.; Oyer, A. J.; Carrillo, J. Y.; Xu, T. O.; Adamson, D. H. Graphene:
Solidification Products International, 2015-2017, Product Development Intern
from interfacial trapping to strong, electrically conductive foams. Proceedings of the AN-
Oxford Performance Materials, 2016-2017, Graduate Researcher - Proxy UConn
TEC 2015 Technical Conference and Exhibition, Orlando, FL, March 23-25, 2015.
ExxonMobile, 2014-2016, Graduate Researcher - Proxy UConn
Kraft, G.M., Hui, T., Bento, J.L., Hire, C.C., Adamson, D.H. Living anionic polymerization: Con-
Corenso, 2014, Consultant
trolling molecular weight, composition and architecture. . Proceedings of the ANTEC 2015
CiDRA 2012-2014, Graduate Researcher – Proxy UConn
Technical Conference and Exhibition, Orlando, FL, March 23-25, 2015
Wisconsin Institute for Sustainable Technology, 2008-2012, Research and Development
Kraft, G. M.; Santiago, A.; Hire, C. C.; Adamson, D. H. Electrospun biomimetic synthetic
Chemist
polymer for templated ceramic condensation. Proceedings of the 248th American Chemi-
cal Society National Meeting, San Francisco, CA, August 10-14, 2014.
HONORS
Droske, J. P.; Juetten, M; Kraft, G. M.; Huberty, W.; Pieper, R. “Green” thermosets: solvent-
less synthesis and reversible crosslinking of poly(alkylene mercaptosuccinates). Polymer
GAANN Fellowship – 2016, 2017
Preprints, ACS Division of Polymer Chemistry 2012, 53 (2), 339.
SPE Connecticut Section Scholarship – 2014, 2015, 2016
Kraft, G. M. Degradation rates of poly(alkylene succinate) analogs under compost condi-
ANTEC 2015 Graduate Poster Competition – 2nd Place
tions. (Grant submitted to) The Student Research Fund, University of Wisconsin-Stevens
Outstanding Student Chapter 2015 – 5th Place Nationally
Point, October 28, 2011.
Doctoral Dissertation Fellowship – Fall 2015
Kraft, G. M.; Sternhagen, G. L.; Droske, J. P. “Green” crosslinkable polymers for sustainable
Predoctoral Fellowship – Fall 2012
applications. Fourth Annual Wisconsin Science and Technology Symposium, University of
SRF Grant 2011
Wisconsin-Whitewater, July 28-29, 2011.
Kraft, G. M. A “green” step-growth synthesis of crosslinkable polyesters. University of Wis-
PUBLICATIONS AND PRESENTATIONS
consin-Stevens Point College of Letters and Science 12th Annual Undergraduate Research
Symposium, April 29, 2011.
Kraft, G.M, Hire, C.C., Santiago, A. Adamson, D.H., “Electrospun biomimetic catalytic poly-
mer template for the sol-gel formation of multidimensional ceramic structures”, Materi-
als Letters, 240:242-245 (2019).
PROFESSIONAL SOCIETY MEMBERSHIPS
Kraft, G. M.; Mohammadi, R.; Adamson, D. H. Controlled synthesis of amphiphilic polymer
American Chemical Society
bottlebrushes. GAANN Fellowship Research Report, U.S. Department of Education, Janu-
Society of Plastic Engineers
ary 20, 2017.
Society of Mineralogical Engineering
Kraft, G. M. Controlled polymer synthesis and 2D sheet based materials. University of
Connecticut Institute of Materials Science Advisory Board Meeting, Storrs, CT, October 26,
032 033Sixth Wave Innovations Inc.
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BRANDI C. MAULL
EDUCATION
M.S. Chemistry, University of Nevada, Reno (2010)
B.S. Biochemistry, Syracuse University (2007)
WORK EXPERIENCE
Sixth Wave Innovations Inc. 2014 to present. Chemist
Raptor Detection Technologies, LLC. June 2012 to 2013. Laboratory Chemist.
Raptor Detection Technologies, LLC. March 2012 to May 2012. Laboratory Technician.
University of Nevada, Reno. August 2002 to May 2010. Graduate Research Associate.
University of Nevada, Reno. August 2002 to May 2010. Teaching Assistant.
Syracuse University. January 2004 to May 2007. Undergraduate Research Associate.
PUBLICATIONS AND PRESENTATIONS
Molecularly Imprinted Polymer Applications for the Gold Mining Industry, G. Southard,
J. Gluckman, B. Maull Proceedings of Heap Leach Solutions, 2015 September 14-16, 2015,
Reno, Nevada, USA.
de Bettencourt-Dias, A., Bauer, S., Viswanathan, S., Maull, B. C. and Ako, A. M., “Unusual
Nitro-Coordination of Europium(III) and Terbium(III) with Pyridinyl Ligands” Dalton Trans- CSE:SIXW | OTC: ATURF | FSE:AHUH
actions, 41,11212-11218 (2012).
Poster Presentation, “Nitro Derivatized Pyridine Derivatives Complexed to Lanthanide
Ions.” American Chemical Society National Meeting (Spring 2009, Salt Lake City, UT).
Sixth Wave Innovations Inc.
PROFESSIONAL SOCIETY MEMBERSHIPS 110-210 Waterfront Drive
Bedford, Nova Scotia, Canada
American Chemical Society (2007)
B4A0H3
Phi Kappa Phi Honor Society (2009)
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