CHEMICAL AND BIOLOGICAL DEFENSE PROGRAMFY17.1: Small Business Innovation Research (SBIR)

Component Specific Proposal Submission Instructions

The approved FY17.1 topics included in the Chemical and Biological Defense (CBD) Small Business Innovation Research (SBIR) Program are listed below. Offerors responding to this Announcement must follow all general instructions provided in the Department of Defense (DoD) SBIR Program Broad Agency Announcement. Specific CBD SBIR requirements that add to or deviate from the DoD SBIR Program Announcement are provided below with references to the appropriate section of the DoD SBIR Announcement.

General Information

In response to Congressional interest in the readiness and effectiveness of U.S. Nuclear, Biological and Chemical (NBC) warfare defenses, Title XVII of the National Defense Authorization Act for Fiscal Year 1994 (Public Law 103-160) required the Department of Defense (DoD) to consolidate management and oversight of the Chemical and Biological Defense (CBD) Program into a single office – Office of the Assistant Secretary of Defense for Nuclear, Chemical and Biological Defense Programs. The Joint Science and Technology Office for Chemical and Biological Defense (JSTO-CBD), Defense Threat Reduction Agency (DTRA) provides the management for the Science and Technology portfolio of the Chemical and Biological Defense Program. Technologies developed under the CBD SBIR Program have the potential to transition to the Joint Program Executive Office for Chemical and Biological Defense (JPEO-CBD) if the appropriate level of technology maturity has been demonstrated. The JSTO-CBD Science & Technology (S&T) programs and initiatives are improving defensive capabilities against Chemical and Biological Weapons of Mass Destruction. The CBD SBIR portion of the CBD S&T Program is managed by the JSTO-CBD and is not part of the DTRA SBIR Program.

The mission of the Chemical and Biological Defense Program is to ensure that the U.S. military has the capability to operate effectively and decisively in the face of chemical or biological warfare threats at home or abroad.  Numerous factors continually influence the program and its technology development priorities.  Improved defensive capabilities are essential in order to mitigate the impact of Chemical and Biological Threats. The U.S. military requires the most up-to-date and state-of-the-art equipment and instrumentation to permit our Warfighters to detect to warn and avoid contamination, if possible, and to be able to sustain operations in a potentially compromised environment.  Further information regarding the DoD Joint Chemical and Biological Defense Program is available at the DoD Counter-proliferation and Chemical Biological Defense homepage at http://www.acq.osd.mil/cp.

The overall objective of the CBD SBIR Program is to identify, obtain and provide technical solutions from small businesses and to improve the transition or transfer of innovative Chem-Bio technologies to the end user – the Warfighter – in addition to commercializing technologies within the private sector for mutual benefit.  The CBD SBIR Program targets those technology efforts that maximize a strong defensive posture in a biological or chemical environment using passive and active means as deterrents.  These technologies include chemical and biological detection for both point and stand-off capabilities; individual and collective protection; hazard mitigation (decontamination); information management, modeling, simulation, and analysis; medical pre-treatments (e.g., vaccine development and delivery); medical diagnostics and disease surveillance; and medical therapeutics (chemical countermeasures and biological countermeasures).

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Submitting Your Phase I CBD SBIR Proposal

The entire SBIR proposal submission (consisting of a Proposal Cover Sheet, the Technical Volume, Cost Volume, and Company Commercialization Report) must be submitted electronically through the DoD SBIR/STTR Proposal Submission system located at https://sbir.defensebusiness.org/.

The Proposal Technical Volume must be no more than 20 pages in length. The Cover Sheet, Cost Volume and Company Commercialization Report do not count against the 20-page Proposal Technical Volume page limit. Pages in excess of this length will not be evaluated and will not be considered for review. The proposal must not contain any type smaller than 10-point font size (except as legend on reduced drawings, but not tables).

If your proposal is selected for award, the technical abstract and discussion of anticipated benefits will be publicly released on the Internet; therefore, do not include proprietary information in these sections. Classified information is never permitted to be included in any SBIR proposal.

Beginning in FY17 (DoD FY17.1 Announcement), the CBD SBIR Program will fund a Phase I proposal up to $150,000 for a six-month period of performance (independent of Discretionary Technical Assistance funding, if requested). The CBD SBIR Program no longer uses a Phase I Option period and will not accept any Phase I proposals that include an option period. Additionally, the CBD SBIR Program will not accept Phase I proposals which exceed $150,000 (exclusive of Discretionary Technical Assistance; see below).

The CBD SBIR Program reserves the right to limit awards under any topic, and only those proposals of superior scientific and technical quality in the judgment of the technical evaluation team will be funded. The offeror must be responsive to the topic requirements, as solicited.

Proposals not conforming to the terms of this Announcement, and any unsolicited proposals, will not be considered. Awards are subject to the availability of funding (subject to the Congressional Budget process) in addition to successful completion of contract negotiations. Note the Chemical and Biological Defense Program is not responsible for any funds expended by the proposer prior to contract award.

CBD Program Phase II Proposal Guidelines

Phase II is the demonstration of the technology that was found feasible in Phase I. Phase I awardees may submit a Phase II proposal without invitation; however, it is strongly encouraged that a Phase II proposal not be submitted until sufficient Phase I progress can be evaluated and assessed based on results of the Phase I proof-of-concept/feasibility study Work Plan and at a recommended five months from date of contract award. All Phase II proposal submissions must be submitted electronically through the DoD SBIR/STTR Proposal Submission system at https://sbir.defensebusiness.org/. At the DoD SBIR Proposal Submission Web site, Phase II proposals MUST be submitted to ‘CBD SBIR’ regardless of which DoD contracting office negotiated the Phase I contract. Additional instructions regarding Phase II proposal submission process including submission key dates will be provided to Phase I awardees after Phase I contract award and also can be found at https://www.cbdsbir.net.

CBD SBIR Phase II Cost Volumes must contain a budget for the entire 24-month Phase II period not to exceed the total maximum dollar amount of $1,000,000. These costs must be submitted using the Cost Volume format (accessible electronically on the DoD SBIR Proposal Submission Web site), and may be presented side-by-side on a single Cost Volume Sheet.  The total proposed amount should be indicated on the Proposal Cover Sheet as the proposed cost with approximately 50% of the total Phase II budget for Year 1 and the balance of the funding budgeted for Year 2.

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The CBD SBIR Program is committed to minimizing the funding gap between Phase I and Phase II activities. The CBD SBIR Program typically funds a cost-plus fixed-fee Phase II award, but may award a firm fixed price contract at the discretion of the Contracting Officer.

Discretionary Technical Assistance In accordance with the Small Business Act (15 U.S.C. 632), the CBD SBIR Program Office will authorize the recipient of a Phase I and/or a Phase II SBIR award to purchase technical assistance services (Discretionary Technical Assistance, DTA), such as access to a network of scientists and engineers engaged in a wide range of technologies, or access to technical and business literature available through on-line data bases, for the purpose of assisting such concerns as:

making better technical decisions concerning such projects;

solving technical problems which arise during the conduct of such projects;

minimizing technical risks associated with such projects; and

developing and commercializing new commercial products and processes resulting from such projects.

If your small business firm is interested in proposing use of a vendor for Discretionary Technical Assistance, you must provide a cost breakdown in the Cost Volume under “Other Direct Costs (ODCs)” and provide a one-page description of the vendor you will use and the specific technical assistance services to be provided. The proposed amount may not exceed $5,000 for Phase I, and $5,000 for each year of a Phase II project. The DTA description should be included as the LAST page of the Technical Volume. This description will not count against the Phase I or Phase II proposal page limit and will NOT be assessed against SBIR proposal evaluation criteria. Approval of technical assistance is not guaranteed and is subject to review of the Contracting Officer.

CBD SBIR Projects Requiring Animal and Human Subjects

Companies should plan carefully for any research involving animal and/or human subjects in addition to the use of any chemical or biological warfare agents. The brief Phase I Period of Performance may preclude plans requiring the use of these materials as well as animal and/or human subjects prior to obtaining all necessary approvals.

The offeror is expressly forbidden to use or subcontract for the use of laboratory animals in any manner without the express written approval of the U.S. Army Medical Research and Material Command's (USAMRMC), Animal Care and Use Review Office (ACURO). Written authorization to begin research under the applicable protocol(s) proposed as part of the CBD SBIR program will be issued after contract award in the form of an approval letter from the USAMRMC ACURO to the recipient. Furthermore, modifications to already approved protocols require approval by ACURO prior to implementation.

Research under CBD SBIR awards involving the use of human subjects, to include the use of human anatomical substances or human data, shall not begin until the DTRA Research Oversight Board (ROB) provides authorization that the research protocol may proceed. Written approval to begin research protocol will be issued from the ROB, under separate notification to the recipient. Written approval from the ROB is also required for any sub-recipient that will use funds obtained from any CBD SBIR awards to conduct research involving human subjects.

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Changes in research involving human subjects shall be conducted in accordance with the protocol submitted to and approved by the ROB. Non-compliance with any provision may result in withholding of funds and or termination of the award.

Key Dates

17.1 Announcement Pre-Release 30 November 201617.1 Announcement Open 10 January 201717.1 Announcement Close 8 February 2017

Phase I Evaluations February – April 2017Phase I Selections No Later Than 7 May 2017Phase I Awards Approx. August 2017

Phase II Proposal Submission Recommend proposal submission no earlier than approximately fivemonths from date of Phase I contract award. Additional instructions regarding Phase II proposal submission process, including key dates, will be provided to Phase I awardees after Phase I contract award and also can be found at https://www.cbdsbir.net/PhaseII.aspx.

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CBD SBIR PROPOSAL CHECKLIST

This is a Checklist of Requirements for your proposal. Please review the checklist carefully to ensure that your proposal meets the CBD SBIR requirements. Failure to meet these requirements will result in your proposal not being evaluated or considered for award.

_____ 1. The Proposal Cover Sheet along with the Technical Volume, Cost Volume, and Company Commercialization Report were submitted via the Internet using the DoD SBIR Proposal Submission Web site at https://sbir.defensebusiness.org/. _____ 2. The proposal cost adheres to the CBD SBIR Program criteria specified.

_____ 3. The proposal is limited to only ONE SBIR topic. All required documentation within the proposal references the same topic number.

_____ 4. The proposal is responsive to the requirements addressed in the topic.

_____ 5. The Project Abstract and other content provided on the Proposal Cover Sheet does not contain any proprietary or classified information and is limited to the space provided.

_____ 6. The Technical Volume of the proposal includes the items identified in Section 5.3.c of the DoD SBIR Program Announcement.

_____ 7. The Proposal Technical Volume must be 20 pages or less in length. The Cover Sheet, Cost Volume and Company Commercialization Report do not count against the 20-page Proposal Technical Volume page limit. Technical Volume pages in excess of 20-pages will not be evaluated and will not be considered for review.

_____ 8. The Company Commercialization Report is submitted online in accordance with Section 5.4.e of the DoD SBIR Program Announcement. This report is required even if the company has not received any prior SBIR funding.

_____ 9. The proposal must not contain any type smaller than 10-point font size (except as legend on reduced drawings, but not tables).

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CBD SBIR 17.1 Topic Index

CBD171-001 Development of Solid-state Optical Cooler Materials to Replace Conventional Cryocoolers Used for Cooling SWIR and LWIR Infrared Detectors and Focal-plane-arrays

CBD171-002 MicroLens Hyperspectral Imager for Standoff Chemical DetectionCBD171-003 Tomographic Nanoscopy for Pathogen IdentificationCBD171-004 Open-Path Optical Analyzer for Quantitative Chemical Weapons DetectionCBD171-005 Process Development and Manufacture of an Antibody-based Therapy Against Sudan Ebola

Virus

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CBD SBIR 17.1 Topic Descriptions

CBD171-001 TITLE: Development of Solid-state Optical Cooler Materials to Replace Conventional Cryocoolers Used for Cooling SWIR and LWIR Infrared Detectors and Focal-plane-arrays

TECHNOLOGY AREA(S): Chemical/Biological Defense, Materials/Processes

OBJECTIVE: To develop efficient Infrared (IR) fluorescent materials for laser/optical cooling to cryogenic temperatures. The materials will serve as cooling agents in the development of all solid-state cryogenic coolers with no moving parts. The goal is to replace current mechanical closed cycle and thermoelectric coolers used in cooling Short Wavelength IR (SWIR) and Long Wavelength IR (LWIR) infrared detectors/Focal-Plane Arrays (FPA). This development will provide remote, lightweight, vibration free cooling and hence more reliable operation of infrared sensor systems designed for surveillance.

DESCRIPTION: There is a need in the Chemical/Biological Defense community for infrared sensors that are both sensitive enough to detect chemical and biological agents, while also being small, lightweight, and inexpensive. In general, cryogenic cooling is required to achieve required sensitivities for chemical and biological detection. Reducing the size, weight and power (SWaP) of current cryogenic systems will provide for improve sensing capabilities. Current LWIR standoff sensors such as the Joint Service Lightweight Standoff Chemical Agent Detector (JSLSCAD) use mechanical closed cycle coolers to achieve cryogenic temperatures needed for operation. Mechanical closed cycle coolers are expensive and unreliable. Much of the expense for the maintenance of JSLCAD units involves the periodic replacement of the mechanical closed cycle coolers. A solid state, no-moving-parts replacement would reduce costs and improve reliability compared to current systems.

The cooling of a solid-state material through anti-Stokes fluorescence may provide an improved method for cooling of infrared detectors [1-4]. Optical cooling of a solid was first demonstrated in 1995 using heavy metal fluoride glass (ZBLANP) doped with Yb3+ ions [1]. The availability of high-brightness laser diodes and fiber-lasers operating in the near infrared region has opened the possibility of practical devices for refrigeration applications based on optical cooling of solids. Advantages of an all solid-state cryocooler include compactness, no vibrations, no moving parts, and no need for cryogenic fluids [1-4]. The current list of solid-state materials, which have shown optical cooling properties, is limited and the net cooling efficiency is relatively low for practical cryogenic applications such as operating IR detector modules. Critical materials requirements for obtaining net radiative cooling from a solid include a near unity emission quantum efficiency, large optical transition cross-sections, no concentration quenching, and low losses due to excited state and unwanted parasitic background absorption. Improvements are being sought in the development of new cooling materials with efficient fluorescence at extended infrared wavelengths (>1 micron) and with improved cooling efficiencies.

The cooling efficiency of a material is affected mainly due to the intrinsic ideal laser-cooling properties of the material, the quality/purity of the material, and the efficient integration of the material into an optical cryocooler device with efficient thermal management. Optical refrigerators based on rare-earth-doped solids have the potential to reach 70 degrees Kelvin (K) and local cooling down to temperatures of ~110 degrees K have been achieved by Seletskiy et al. [5]. Ideal laser cooling efficiencies employing Yb3+ doped materials operating at ~1 micron are limited to ~2-3%. Extending the fluorescence to longer IR wavelengths provides the potential for significantly higher cooling efficiencies [3]. For examples Tm3+ doped solids fluorescence at ~2 microns have the potential to exhibit cooling efficiencies of ~4-5%. The highest cooling efficiencies of ~6-10% can be achieved from materials doped with Dy3+ or Nd3+ ions, which can produce efficient 3-5 micron fluorescence when doped into halide crystals with low maximum phonon energies (< 300 wavenumbers) [6,7]. The improvements in mid-IR laser sources such as fiber lasers, Cr2+-ZnSe tunable lasers, optical parametric oscillators, and quantum cascade lasers provide new possibilities for efficient coolers operating at longer IR wavelength (2-5 microns). Efficient optical refrigeration is only possible if materials with suitable purity and optimal intentional rare earth doping are developed.

PHASE I: Examine key parameters for IR fluorescent materials that can be used in the development of new laser cooling devices using anti-Stokes fluorescence. Examine and identify potential materials that can be used for laser

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cooling using infrared laser radiation at longer wavelengths. Using key parameters, design a new material and develop the pathway to achieve bulk cooling, at least down to 110 degrees K. Demonstration to such level of cooling is desirable.

PHASE II: Develop optical cooling material preparation methodology to achieve efficient cooling. Critical issues limiting the attainable cooling efficiencies in selected optical materials including absorption losses (e.g. excited state absorption, other impurities). Re-absorption and extraction efficiency must be studied and controlled. The Phase II program must demonstrate reproducible production of the material to achieve the limit of cryogenic cooling temperatures and provide a road map to achieve liquid nitrogen temperatures (77 degrees K). Demonstration of 77 degrees K temperature is desirable. Integrating the new material in a form of a practical device, capable of handling thermal load equivalent to typical single element, linear array and FPA, should be explored. Thermal issues involved in device integration should be identified, analyzed, and controlled. The Phase II deliverables shall include a final report detailing the production of cooling material, cooling device design and sample optical cooling materials as well as a prototype integrated optical cooling device.

PHASE III DUAL USE APPLICATIONS: PHASE III: Further research and development during Phase III efforts will be directed toward refining and implementing the new cryocooler to meet the requirements of the Joint Chemical and Biological Defense Program in addition to U.S. Army Concept of Operations (CONOPS) and end-user requirements. The new cooler will have broad impact across several avenues of defense applications. Engineering and prototype development, test and evaluation, and hardware qualification demonstration in a system-level test-bed, which shows application to an insertion potential into one or more SWIR and LWIR FPAs, will be examined.

Possible commercial applications include satellite based and space based remote sensor system cooling of electronic systems and optical refrigeration. Cyrogenic cooling will be used in commercial sensing applications, including industrial applications.

REFERENCES:1. R. I. Epstein, M. I. Buchwald, B. C. Edwards, T. R. Gosnell, and C. E. Mungan, “Observation of laser-induced fluorescent cooling of a solid,” Nature 377, 500-503 (1995).

2. Markus P. Hehlen, Proc. of SPIE Vol. 7228 72280E-1 (2009).

3. C.W. Hoyt, M. P. Hasselbeck, M. Sheik-Bahae, R. I. Epstein, S. Greenfield, J. Thiede, J. Distel, and J. Valencia, J. Opt. Soc. Am. B, Vol. 20, 1066 (2003).

4. Denis V. Seletskiy, Seth D. Melgaard, Stefano Bigotta, Alberto Di Lieto, Mauro Tonell & Mansoor Sheik-Bahae, Nature Photonics 4, 161 - 164 (2010).

5. Denis V. Seletskiy, Seth D. Melgaard, Richard I. Epstein, Alberto Di Lieto, Mauro Tonelli, and Mansoor Sheik-Bahae; Optics Express, 19, 2011, 18229.

6. E. Virey, M. Couchaud, C. Faure, B. Ferrand, C. Wyon, C. Borel, “Room temperature fluorescence of CsCdBr3: Re (Re=Pr,Nd,Dy,Ho,Er,Tm) in the 3-5µm range”. J. Alloys and Compounds 257-277, (1998) 311.

7. U. Hömmerich, E.E. Nyein, P. Amedzake, S. B. Trivedi, J.M. Zavada, “Mid-Infrared (3-5µm) emission from rare earth doped KPb2Br5”, Proceedings of SPIE Vol. 6100, (2006) 61000G-18.

KEYWORDS: Optical Cooling Materials, Infrared Detectors, anti-Stokes fluorescence, cryogenic cooling; cryocooler

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CBD171-002 TITLE: MicroLens Hyperspectral Imager for Standoff Chemical Detection

TECHNOLOGY AREA(S): Chemical/Biological Defense

OBJECTIVE: Develop a small lightweight lenslet-based hyperspectral imaging sensor for detection and identification of chemical agents and toxic industrial chemicals

DESCRIPTION: Hyperspectral imaging (HSI) sensors currently in use for wide-area standoff detection of chemicals are required to interrogate a large number of spectral bands in order to differentiate between target compounds and the background environment. The sensors typically required a large focal-plane-array to achieve necessary spatial coverage and spatial resolution for wide area chemical detection. However, a chemical standoff sensor also needs to be small, lightweight, and inexpensive. The conflicting requirements of high performance in a small, inexpensive package provide challenges for future development. The goal of this topic is to reduce the size, weight, power, cost, and bandwidth of current HSI systems while maintaining the capability of wide area early warning of a chemical incident.

Microlenses are small lenses with diameters less than one millimeter. Often microlenses are fabricated as small as 10 micrometers. Since the lenses are extremely small, good optical quality can be achieved with a very simple design. A typical microlens may be a single element with one plane surface and one spherical convex surface to refract the light. More sophisticated lenses may use aspherical surfaces. Others designs use several layers of optical material to achieve good optical quality. Another type of microlens utilizes two flat and parallel surfaces with the focusing action achieved by a variation of refractive index across the lens. Such lenses are known as gradient-index (GRIN) lenses. Some microlenses achieve their focusing action utilizing both a variation in refractive index and surface shape.

A properly designed lenslet or microlens arrays can also be used to extract simultaneous spectral and spatial information from a hyperspectral scene. Thus, microlenses can be utilized in the fabrication of extremely small imaging spectrometers. The goal of this effort is the development of micro-machined structures for miniature diffractive optical elements that are not currently achievable. This research will examine programmable electronics that could be integrated with focal plane arrays. The tight integration of focal-plane-arrays with programmable electronics could ultimately allow for the development of a monolithic focal plane array with integrated drive and processing electronics that would provide real time spectral image processing. The ultimate goal of this effort is to design and build an autonomous miniature imaging spectrometer.

PHASE I: Determine the parameters for a lenslet array that would be required in the development of an infrared hyperspectral imager. Emphasis should be placed on the thermal infrared region of the spectrum (8 to 12 μm). Examine available infrared focal plane array technologies and monitor expected advancements in this area in the near future. Conduct a first order lenslet mosaic array design and examine processes for the fabrication of this design. The lenslet design should provide both spectral and spatial information to the ultimate user of the system. A sensitivity model and prediction based on practical components should be completed. The goal of this effort is to develop an imaging standoff chemical detector system that weighs less than five (5) pounds in an overall package of less than 50 cubic inches that is able to operate using battery power (the 5-pound weight objective includes the weight of the batteries). The system should also demonstrate significant cost reduction as compared to traditional wide area standoff chemical detection systems that utilize hyperspectral imaging.

PHASE II: Fabricate a lenslet-based hyperspectral imaging system operating in the thermal infrared region of the spectrum (8 to 12 μm). Design and build all necessary readout electronics. Test and characterize the system. Use the results of the testing to update/improve the sensor design, where identified.

PHASE III DUAL USE APPLICATIONS: Further research and development during Phase III efforts will be directed towards a final deployable design, incorporating design modifications based on results from tests conducted during Phase II, and improving engineering/form-factors, equipment hardening, and manufacturability designs to meet the requirements of the Joint Chemical and Biological Defense Program in addition to U.S. Army Concept of Operations (CONOPS) and end-user requirements. Further, demonstrate the technology’s applicability to stand-off

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detection of chemical and biological threat materials.

There are environmental applications for a small robust, chemical sensor. A rugged, low-cost chemical sensor will benefit the manufacturing community by providing inexpensive monitoring of chemical processes. Also, first responders such as Civil Support Teams and Fire Departments have a critical need for a rugged, inexpensive sensor that can be transported to the field to test for possible contamination by chemical agents and toxic industrial chemicals.

REFERENCES:1. Bogdan R. Cosofret, Shin Chang, Michael L. Finson, Christopher M. Gittins, Tracy E. Janov, Daisei Konno, William J. Marinelli, Mark J. Levreault, and Rex K. Miyashiro, “AIRIS standoff multispectral sensor”, Proceedings of the SPIE, volume 7304, pages 73040Y (2009).

2. Vincent Farley, Charles Belzile, Martin Chamberland, Jean-Francois Legault, and Karl R Schwantes, “Development and testing of a hyperspectral imaging instrument for field spectroscopy”, Proceedings of SPIE, volume 5546, pages 29-36 (2004).

3. Ju-Seog Jang and Bahram Javidi, “Improved viewing resolution of three-dimensional integral imaging by use of nonstationary micro-optics”, Optics Letters, Vol. 27, Issue 5, pages 324-326 (2002).

4. E. Tajahuerce, P. Andrés, J. Lancis, M. Fernández-Alonso, and V. Climent, “White-light array generation with a diffractive lenslet array”, Journal of Modern Optics, vol. 46, Issue 1, pages 49-63 (1999).

5. Shijun Xiao, A.M. Weiner, and C. Lin, “A dispersion law for virtually imaged phased-array spectral dispersers based on paraxial wave theory”, IEEE Journal of Quantum Electronics, Volume 40, Issue 4, pages 420- 426 (2004).

6. Dan Daly, “Microlens Arrays”, CRC Press (2001).

7. D.A. Scribner, M.R. Kruer, and J.M. Killiany, “Infrared focal plane array technology”, Proceedings of the IEEE, Volume 79, Issue 1, pages, 66-85 (1991).

8. Michele Hinnrichs and Mark A. Massie, “New approach to imaging spectroscopy using diffractive optics”, Proc. SPIE, Vol. 3118, pages 194-205 (1997).

9. M. Hinnrichs, N. Gupta, A. Goldberg, “Dual band (MWIR/LWIR) hyperspectral imager”, Proceedings of the 32nd Applied Imagery Pattern Recognition Workshop, pages 73- 78 (2003).

KEYWORDS: standoff chemical detection, hyperspectral imaging, MEMS, lenslet array, microlens, infrared spectrum

CBD171-003 TITLE: Tomographic Nanoscopy for Pathogen Identification

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: Develop an optical method for the determination of cellular structure and function in possible pathogen organisms

DESCRIPTION: Label- and reagent-free techniques for three-dimensional (3D) cell imaging with tens of nanometer spatial resolution are needed to characterize the structure and function of potential pathogen organisms. The approach should provide specific biomolecular functional information which will further aid in discrimination. A

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real time capability would further enhance the utility for time-resolved observation of the organism as it reacts to environmental modifications or stimuli and therapeutic or antibiotic activity.

Historically, sub-diffraction scale resolution has been obtained through fluorescence saturation techniques, typically through manipulating the excitation intensity through interference. These techniques have been limited to certain fluorescent proteins and do not yield functional or 3D information. Nanometer scale molecular confirmation determination has been performed using a 4π configuration and spectral self-interference relative to a reflective surface. This technique on the other hand is not label-free. Recently, high resolution (label-free) phase analysis has become of interest for morphological and composition determination of a specimen in areas of tissue disease diagnosis. Diffraction limited phase measurements are possible using digital holographic microscopy (DHM) which is ideal for complex electromagnetic wave fields scattered by biological tissue. Recent efforts have employed DHM in a quasi-2π configuration with violet illumination and reported unprecedented lateral resolution (60 nm). Moreover, by scanning a synthetic aperture and recording a series of projections through the sample, tomographic 3D reconstruction can be accomplished.

There is a need to augment current tomographic nanoscopy methods to develop better detection and diagnostic capabilities. A functional capability, such as Raman or infrared absorption spectroscopy, would support species discrimination based on the biomolecular make-up of the organism. High-speed data generation and analysis would enable time-resolved measurements of the organism’s response to environmental changes or to therapeutics such as antibiotics. These enhanced capabilities will promote better discrimination of the organism and a more intrinsic understanding of the organism’s behavior as it relates to treatment discovery.

PHASE I: Generate a preliminary design of a tomographic nanoscopic cell imaging system capable of biomolecular functional determination. Perform a system level analysis predicting the overall performance including key parameters such as lateral and axial resolutions, sensitivity, specificity, and baseline computational timing. Generate an opto-mechanical design and identify the key components. Breadboard subsystems identified to be high in technical risk. Generate a preliminary data processing and reconstruction design.

PHASE II: Design, build, and test a prototype tomographic nanoscale cell imaging system. Optimize the processing and reconstruction algorithms for near real time. Fully characterize the system. Demonstrate a series of relevant cellular measurements, including a time-resolved series of a cell’s response to external stimuli. Deliver the prototype system to the government.

PHASE III DUAL USE APPLICATIONS: During Phase III, refine a final design tailored for a specific application or platform, incorporating design modifications based on results from tests conducted during Phase II, and improving engineering/form-factors, processing/reconstruction, and manufacturability designs to meet requirements of the Joint Chemical and Biological Defense Program and end-user requirements.

This type of technology will also find use in medical research laboratories for cell and tissue diagnostics, biomarker discovery and identification and in pharmaceutical laboratories for drug discovery.

REFERENCES:1. M.G.L. Gustafsson, “Surpassing the lateral resolution limit by a factor of two using structured illumination microscopy,” J. Microsc. 198, 82-87 (2000).

2. B.J. Davis,1, M. Dogan, B.B. Goldberg, W.C. Karl, M. S. Ünlü, and A.K. Swan, “4Pi spectral self-interference microscopy,” JOSA A 24(12), 3762-3771 (2007).

3. Y. Cotte, F. Toy, P, Jourdain, N. Pavillon, D. Boss, P. Magistretti, P. Marquet, and C. Depeursing, “Marker-free phase nanoscopy,” Nat. Photonics 7, 113-117 (2013).

4. E. Cuche, P. Marquet, and C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl. Opt. 38(34), 6994-7000 (1999).

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5. P. Marquest, et al., “Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with sub-wavelength accuracy,” Opt. Lett. 30, 468 (2005).

6. D. Naumann, et al., “The characterization of microorganisms by Fourier Transform Infrared Spectroscopy (FTIR),” in Modern Techniques for Rapid Microbiological Analysis, W.H. Nelson, ED., VCH Publishers, New York, 1991.

7. D. Whelan, et al., “Monitoring the reversible B to A-like transition of DNA in eukaryotic cells using Fourier transform infrared spectroscopy, Nucleic Acids Research 39, 5439 (2011).

KEYWORDS: nanoscopy, spectroscopy, tomography, cell imaging, pathogen identification, sub-diffraction scale

CBD171-004 TITLE: Open-Path Optical Analyzer for Quantitative Chemical Weapons Detection

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: Develop inexpensive dynamic, real time analytical technology for multicomponent vapor identification and quantification.

DESCRIPTION: Chemical detection technologies rely on intrinsic molecular signatures of the targeted threat agents present in a sampled space vapor or aerosol form. Detection of the signature is interpreted as an actionable event without specific regard to the quantitation of the suspected threat agent concentration. As advances in network architectures and decision logic mature, there is increasingly a need to quantify the concentration of a suspected threat plume in order to address the likelihood of a true positive detection event and to characterize the nature and extent of contamination or hazard presented by the incident. Recent developments in optical analysis techniques may afford alternatives to extant mass-based methods (ion mobility spectrometry or mass spectrometry), affording the potential for an open-path construct for analyzing a small volume for both the presence of a threat analyte and its instantaneous concentration while obviating the need for volumetric sampling of the tested volume through an inlet using an active airflow. Such an architecture would have the advantage of rapid analysis over several orders of magnitude in threat species concentration. The threat agents of concern include the organophosphate and organosulphur compounds that typify the chemical agent threat space.

PHASE I: The Phase I feasibility/proof-of-concept study will thoroughly analyze and model the component and system level performance of an optical analyzer based on laser/dynamic photoacoustic, and/or cavity ringdown spectroscopy in order to develop an end-to-end theoretical performance model and to drive the preliminary design constraints for a Phase II demonstration system. Key performance parameters associated with the technical approach must include detection and quantification of airborne chemical threats to include chemical warfare agents (CWAs) and toxic industrial chemicals (TICs). The analytical approach should at a minimum classify these compounds at the 10 minute Negligible Military Exposure Guideline (MEG) concentrations (see TG-230), with sufficient discrimination against background interferents, in order to indicate a hazardous environment. The signal-to-noise characteristics and dynamic range of the analytical approach must be modeled to demonstrate suitable sensitivity and linearity across five decades of concentration levels to chemometrically quantify the following CWAs and TICs:• CWA vapors of chemical purity greater than 40%: analytes of interest include GA, GB, GD, GF, HD, HN3, Lewisite (required) and VX (optional)• TIC vapors of chemical purity greater than 80%: analytes of interest include at minimum hydrogen cyanide (HCN), cyanogen chloride (ClCN), phosgene (COCl2), hydrogen sulfide (H2S), ammonia (NH3), Nitrogen Dioxide (NO2), and Sulfur Dioxide (SO2)

The Edgewood Chemical Biological Center (ECBC) quantitative infrared spectral database will be made available to offerors by request. Additional data are accessible in the Pacific Northwest National Laboratory (PNNL)

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quantitative infrared spectral database.

Affordability, response time, and size, weight, and power (SWaP) is a critical evaluation criterion for the candidate technology. In order to be competitive and suitable for military threat monitoring, the technology cannot cost in excess of $10,000 per system in production (assume a scale of 100s of units per year), must respond in 60 seconds or less, and must not exceed the following size, weight, and power constraints:• Must weigh less than one (1) kilogram (kg) including installed batteries and be no larger than 650 cubic centimeters (cm3)• Must Operate for 24 hrs. using standard commercial off-the-shelf (COTS) batteries.

PHASE II: The Phase II effort will fabricate, integrate, test, and optimize the performance of a real-time open-path vapor analysis prototype platform based on the outcome of the Phase I feasibility study. Offerors should arrange quantitative agent assessment tests of the prototype platform with a laboratory authorized to handle and manipulate the various CWAs and TICs. The prototype system cannot be recovered from the chemical surety laboratory after exposure to CWAs.

PHASE III DUAL USE APPLICATIONS: During Phase III, a real time, quantitative, multicomponent analytical capability will enable the development of novel air-monitoring technology suitable for application in a variety of defense and security settings as well as industrial and commercial environments.

The capability to quantitatively define the concentration of gas phase constituents in real time would lead to new products for process and environmental quality monitoring in the pharmaceutical, semiconductor, and advanced materials industries.

Additionally, the quantitative vapor/aerosol monitoring technology developed under this effort would enable a wide variety of application models, to include compliance, safety and health, medicinal/diagnostic monitoring, and industrial/medical process monitoring.

Offerors should enunciate a clearly-defined commercialization strategy for the gas monitoring and quantification technology that includes an analysis of the market for these and other safety and health, environmental surveillance, diagnostic, and industrial monitoring applications.

REFERENCES:1. TG-230 Environmental Health Risk Assessment and Chemical Exposure Guidelines for Deployed Military Personnel: http://phc.amedd.army.mil/PHC%20Resource%20Library/TG230.pdf

2. ECBC Spectral Database: https://cbconnect.apgea.army.mil/RT/ECBCSpectraDB/default.aspx. Requestors must also request an Army Knowledge Online (AKO) account in order to access the SharePoint. Requests for temporary AKO accounts will be supported by this topic’s author: https://login.us.army.mil/suite/login/

3. Johnson TJ, RL Sams, and SW Sharpe. 2004. "The PNNL Quantitative Infrared Database for Gas-Phase Sensing: A Spectral Library for Environmental, Hazmat and Public Safety Standoff Detection." In Chemical and Biological Point Sensors for Homeland Defense, vol. 5269, ed. AJ Sedlacek III, R Colton and T Vo-Dinh, pp. 159-167. SPIE, Bellingham, WA. doi:10.1117/12.515604

4. A.A. Kosterev, P.R. Buerki, L. Dong, M. Reed, T. Day, F.K. Tittel, "QEPAS detector for rapid spectral measurements", Appl Phys B, 101, 649 (2010)

5. Lei Dong, Frank K Tittel, Chunguang Li, Nancy P Sanchez, Hongpeng Wu, Chuantao Zheng, Yajun Yu, Angelo Sampaolo, Robert J Griffin, "Compact TDLAS based sensor design using interband cascade lasers for mid-IR trace gas sensing", Opt Express 24(6) A528-A535 (2016)

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6. Wei Ren, Wenzhe Jiang, and Frank K. Tittel, "QCL Based Absorption Sensor for Simultaneous Trace-Gas Detecton of CH4 and N2O" Appl Phys B 117(1), 245-251 (2014)

7. C. M. Wynn, S. Palmacci, M. L. Clark, and R. R. Kunz, “High-sensitivity detection of trace gases using dynamic photoacoustic spectroscopy,” Opt. Eng. 53(2), 021103 (2014)

8. R. M. Sullenberger, M. L. Clark, R. R. Kunz, A. C. Samuels, D. K. Emge, M.W. Ellzy, and C. M. Wynn, "Trace aerosol detection and identification by dynamic photoacoustic spectroscopy", Opt Express 22(S7) A1810-A1817 (2014)

KEYWORDS: Cavity Ringdown Spectroscopy, Dynamic Photoacoustic Spectroscopy, Quartz-Enhanced Photoacoustic Spectroscopy, Chemical Agent, Open Path Infrared, Quantitative Spectroscopy

CBD171-005 TITLE: Process Development and Manufacture of an Antibody-based Therapy Against Sudan Ebola Virus

TECHNOLOGY AREA(S): Biomedical, Chemical/Biological Defense

OBJECTIVE: Develop and optimize a bioprocess for the clinical manufacture of an antibody-based therapy against Sudan ebolavirus

DESCRIPTION: Filoviruses continue to pose a significant bioterrorism threat due to their high mortality rate, transmissibility, and the lack of an approved therapy or vaccine. The National Institute of Allergy and Infectious Diseases (NIAID) Category A biodefense pathogens Marburg virus (MARV) and Ebola virus (EBOV) belong to the Filoviridae family of enveloped negative-sense RNA viruses [1]. Zaire and Sudan viruses (SUDV) are responsible for the largest number of human Ebola virus cases and deaths to date and represent the chief Ebola virus species for which vaccines and therapeutics are currently being developed [2]. Fatality rates associated with infection can be as high as 90% in humans [3]. Recent successes by several independent groups have provided convincing evidence that antibody-based therapies can be an effective countermeasure against filovirus infection. Dye et al. were the first to demonstrate protection of macaques against Ebola and Marburg virus infection using antibody-based immunotherapies [4]. In these studies, EBOV or MARV infected naïve rhesus macaques were passively transferred fractionated EBOV or MARV-specific IgG purified from convalescent Non-Human Primates (NHPs) provided 100% protection against EBOV and MARV challenge even when initiation of treatment was delayed until 48 hours post infection. However, transfer of this concept to a Food and Drug Administration (FDA) approved human-use product is impractical because the antibody immunotherapy source is convalescent serum isolated from filovirus survivors. Therefore, the development and testing of a non-BioSafety Level-4 (BSL-4) reagent is essential for the development of a safe and reproducible therapeutic product. A valuable lesson learned during the large Zaire Ebola virus outbreak of 2014-15 in West Africa is the lack of scalable manufacturing capability for pipeline products available for Emergency Use Authorization (EUA). Case in point was the existing stock of 5-7 total doses of ZMapp, an antibody cocktail against Zaire virus, that was immediately exhausted [5]. Scale-up of the plant-based manufacturing process for rapid delivery of ZMapp to those in need was not possible, in part, as such surge capacity for plant production had not been contemplated in the design of the U.S. Government’s Centers for Innovation in Advanced Development and Manufacturing (CIADM).

While several mAb cocktails specific for Zaire virus have been developed and tested, development efforts for Sudan virus-specific antibody immunotherapy is currently lacking. There exists a tremendous void in the Ebola virus therapeutic arsenal in terms of Sudan virus-specific countermeasures, leaving the U.S. vulnerable to Sudan virus infection with no way of actively treating or managing outbreaks. Previous and ongoing monoclonal antibody development efforts led by the Dye group at the U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID) have yielded a well characterized panel of murine-derived and primate single chain variable fragment (scFv)-derived antibodies specific for SUDV GP. These mAbs have been downselected to a group of lead candidates for therapeutic development. The goal of this topic is to develop, optimize and scale-up a mammalian cell culture-

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based bioprocess for the rapid manufacture of an antibody-based immunotherapy against Sudan virus. The selected awardee must provide a mammalian cell culture based-process capable of rapid scale-up and translation to the CIADM facilities currently existing under the Biomedical Advanced Research and Development Authority (BARDA) and those under construction by the Department of Defense Medical Countermeasures Advanced Development and Manufacturing Capabilities (MCM ADMC). The bioprocess must be fully compatible with single-use bioreactor (SUB) systems, rapidly scalable for surge requirements, and highly productive (greater than 1 gram per liter for all mAb components). Innovative improvement of FDA-approved cell lines for significantly increasing specific productivity are encouraged. Optimization of cell line generation and production medium to eliminate animal serum, minimize animal derived components, and translate the bioprocess to current Good Manufacturing Practices (cGMP) single-use manufacturing is required. The selected awardee should have extensive experience in the construction and expression of mAbs, particularly antiviral therapeutics, and should have the resources needed for product and process characterization. The successful applicant would also need to consider and demonstrate knowledge of all relevant manufacturing issues that are required in therapeutic mAb development – these include the adherence to cGMP guidelines, the historical requirements for FDA approval of such products under the Animal Rule, and the economic feasibility of production. This topic supports EO 13329 requiring SBIR/STTR agencies to give high priority to manufacturing-related research and development.

PHASE I: Generate suspension adapted mammalian cell lines with high specific productivities (target of 30 picograms per cell per day or greater). Approaches may include combinations of multiple selection strategies and innovative means for attaining higher transgene copy number, and must be animal serum free. Preferably, the parental cell line will be readily acceptable by FDA (i.e., established track record for mAb manufacture). At least two mAb producing cell lines will be required for the envisioned therapeutic cocktail. Perform rapid cloning of each cell line to select lead and backup production cell line for each mAb component. Approaches may include robotic automation and high resolution imaging integrating elements of high throughput screening for reduced timelines and high confidence of clonality. Provide at least 50 mg of purified mAb for small animal evaluation for each lead and backup clonal cell line.

PHASE II: Define a Target Product Profile (TPP) for each mAb component. Characterization may include high resolution chromatography, mass spectrometry or other means to establish Critical Quality Attributes (CQAs) that may affect safety and potency of the therapeutic. Develop bioprocesses for cGMP manufacture of the two mAb components. Unit operations must be fully transferrable to single-use systems for rapid scale-up and surge production. Upstream bioreactor processes must be fully defined with no animal derived components. Approaches may include fed-batch or perfusion modes to attain multigram per liter volumetric yields. Application of scalable single-use technologies for harvest and purification such as continuous processing is encouraged. Demonstrate stability of each lead clonal cell line. By the end of Phase II/Year 1, produce each component at small scale (10-50L) for generation of preclinical materials. Characterize both bioprocess and product to establish in-process and release assays. Validate assays as needed for Phase I cGMP manufacturing. Perform GLP engineering runs at a representative scale (>50L) for production of IND-enabling studies and to demonstrate scalability for transfer of processes to cGMP for Phase I clinical trials. Deliver bulk drug substance (BDS) and related Certificates of Analysis (CoA) and Batch Production Records (BPRs) to support pre-IND meeting with the FDA.

PHASE III: The objective of continuing this project into SBIR Phase III is to advance the Sudan Ebola virus therapeutic into the clinic and through FDA licensure. The offeror will advance mAb bioprocesses to cGMP production scale suitable for FDA allowed Phase I clinical investigations. Perform viral clearance studies to assure sufficient virus reduction in downstream processes. Transfer, translate and/or scale-up processes for Phase I cGMP manufacture of BDS to support both human safety and pivotal animal efficacy trials.

PHASE III DUAL USE APPLICATIONS: Project success will result in innovative commercial biomanufacturing technologies inserted into the biodefense system and will also include potential dual use applications of such technologies to non-defense pharmaceutical production.

REFERENCES:1. Fields BN, Knipe DM, Howley PM (2007) Fields virology. Philadelphia: Wolters Kluwer Health/Lippincott Williams & Wilkins. 2 v. (xix, pp. 3091, 3086).

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2. http://www.cdc.gov/ncidod/dvrd/spb/mnpages/dispages/ebola.htm

3. Sanchez A. GTW, Feldmann H. (2007) Filoviridae: Marburg and Ebola viruses. In: Knipe DM HP, Griffin DE, Lamb RA, Martin MA, Roizman B, Straus SE, editor. Fields' Virology. Fifth ed. Philadelphia, PA: Lippincott Williams and Wilkins. pp. 1409-1448.

4. Dye JM, Herbert AS, Kuehne AI, Barth JF, Muhammad MA, et al. (2012) Postexposure antibody prophylaxis protects nonhuman primates from filovirus disease. Proc Natl Acad Sci U S A 109: 5034-5039.

5. Department of Defense, Chemical and Biological Defense Program. 2015 Annual Report to Congress. http://www.acq.osd.mil/cp/cbd_docs/reports/2015_DoD_CBDP_ARC_Final.pdf

KEYWORDS: Ebola; biomanufacturing; monoclonal antibody; cGMP; filovirus; biological agent; medical countermeasure

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