OUSD (R&E) CRITICAL TECHNOLOGY AREA(S): Military Infectious Diseases
OBJECTIVE: Develop and validate a high-throughput psoralen/ultraviolet A(UV-A) based pathogen inactivation device capable of inactivating pathogens at a wide range of volumes (from 0.01 L to 50 L). The solution can facilitate rapid development of vaccines against any emerging infectious threats to protect civilian and military personnel against infectious diseases and reduce lost duty days.
DESCRIPTION: Emerging pathogens with epidemic and pandemic potential are a significant threat to US Forces that defend the homeland and US interests abroad. Historically, highly pathogenic novel viruses have impacted continuity of operations of US Forces with grave consequences. Recently, the COVID-19 pandemic has impacted operations, training, and military readiness across all services and has introduced quarantine and isolation challenges to the US Fleet carrying out freedom of navigation operations in the Pacific. It is imperative to develop, validate, and field an agile vaccine platform that can be rapidly adapted to produce a preventative countermeasure for the next emerging disease threat to US forces. To that end Naval Medical Research Command (NMRC) has developed a psoralen/UV-A based whole virus inactivation method in laboratory scale [1, 2]. NMRC has also developed and optimized a two-step chromatographic method to obtain highly purified psoralen inactivated whole virus vaccine candidates in large quantities to conduct preclinical immunogenicity and efficacy evaluations. We have prepared highly purified monovalent and tetravalent psoralen-inactivated dengue virus vaccines (DENV PsIVs) and a psoralen-inactivated SARS-CoV-2 (SARS-CoV-2 PsIV) vaccine candidate and evaluated their immunogenicity, efficacy and safety in animal models [3, 4]. Psoralen-UV-A based inactivation method can be easily adapted to develop whole cell inactivated vaccines against any pathogen including bacteria, viruses, and parasites, and has a great potential as an agile vaccine platform to rapidly develop vaccines against emerging infectious threats. Based on the preclinical immunogenicity data NMRC is currently working on establishing a contract with a commercial manufacturing organization to make SARS-CoV-2 PsIV under cGMP conditions to conduct a first in human Phase 1 clinical trial. However, our efforts to manufacture the GMP product is hampered by lack of a suitable psoralen/UV-A inactivation device for effectively inactivating pathogens at large enough volumes (10 mL – 50 L batches) under cGMP conditions. The prototype device developed in this SBIR topic should have the capability to inactivate pathogens in 10 mL to 50 L volume using psoralen and UV-A irradiation and the ability to adjust and optimize the parameters such as flow rate of the pathogen solution into and out of the UV-A irradiation chamber, time of UV-A irradiation and the total UV-A energy applied to the pathogen solution for achieving a complete pathogen inactivation without degrading the antigenic proteins. Availability of such a high-throughput psoralen-inactivation device for manufacturing the GMP product will significantly advance the psoralen-inactivated whole cell vaccine platform as an agile vaccine platform against emerging infectious diseases. This pathogen inactivation device can also be used for rapid inactivation of pathogens requiring high containment (BSL-3 and BSL-4 laboratories) without degrading their surface proteins and antigens before bringing them out of the BSL-3 or BSL-4 lab for antigen discovery and characterization.
PHASE I: The main goal of Phase I is a feasibility study towards developing a prototype high throughput UV-A irradiation device capable of handling 10 mL to 50 L volume of pathogens at high titers (greater than 1011 PFU or CFU per mL) while uniformly delivering the UV-A energy to the pathogen solution to achieve complete inactivation of the pathogen. NMRC Tech Transfer office and NMRC legal will work with the small business to license or otherwise distribute prior technology findings from NMRC to awardees at no cost. The proposed psoralen-UV-A irradiation should be a flow through inactivation device with inlets for pumping psoralen/pathogen mixtures into to the UV-A-irradiation chamber with a control the flow rate, and capable of uniformly delivering the UV-A energy to the entire psoralen/pathogen mixture as it flows within the UV-A chamber, and an outlet from the UV-A irradiation chamber to collect the psoralen/UV-A processed inactivated pathogen into an appropriate bioprocessing container for downstream vaccine development processes. The prototype device should include the software and control switches necessary to regulate/adjust all the parameters including flow rate, stop and start flow, amount of UV-A energy applied (microjoules/second/cm2), and the time of application of UV-A energy. The design should include selection of tubing and materials that are low binding to ensure minimal loss of biological material during the inactivation process. Device design should allow for adjusting the total inactivation volumes of pathogens as required.
Phase I deliverables:
- Data demonstrating flow-through inactivation of 10 mL – 5 L of a virus, using psoralen-UV-A irradiation-based pathogen inactivation method/device.
PHASE II: The main objective of Phase II is to develop and produce a fully functional prototype high throughput UV-A irradiation device that is capable of handling 10 mL to 50 L volume of pathogens at high titers (greater than 1011 PFU or CFU per mL) while uniformly delivering the UV-A energy to the pathogen solution to achieve complete inactivation of the pathogen. The major components of the device should include a) an inlet to add specific amount of psoralen derivative to the entire volume of the pathogen, b) an inlet to the UV-A-irradiation chamber and a pump to control the flow rate, c) the UV-A irradiation chamber capable of uniformly delivering the UV-A energy to the entire pathogen solution contained within the UV-A chamber, d) control switch to regulate/adjust the amount of UV-A energy applied to the pathogen solution, and an outlet from the UV-A irradiation chamber to collect the psoralen/UV-A processed inactivated pathogen into an appropriate bioprocessing container for downstream vaccine development processes. The prototype device should include the software and control switches necessary to regulate/adjust all the parameters including flow rate, stop and start flow, UV-A energy applied (microjoules/second/cm2), and the time of application of UV-A energy.
Phase II deliverables:
- One fully functional prototype psoralen-UV-A irradiation-based pathogen inactivation device with data demonstrating complete inactivation of 50 L of Dengue virus.
PHASE III DUAL USE APPLICATIONS: The main target of this high throughput psoralen/UV-A irradiation-based pathogen inactivation device is the GMP vaccine manufacturers who will be making the whole cell inactivated vaccines. Whole virus inactivated vaccines occupy a large proportion of the global viral vaccines market since they elicit a broad range of immune responses and have several advantages including safety and relatively low production cost. The global inactivated vaccines market is expected to increase by more than 10% from 2022 to 2027 (https://www.globalmarketestimates.com/market-report/inactivated-vaccine-market-3754). Formaldehyde and β-propiolactone, the chemicals currently used for making whole virus inactivated vaccines, are less than optimal since they alter the immunogenic proteins and are considered carcinogenic. Psoralen compounds on the other hand do not affect the immunogenic proteins and have been shown to be safe for use in biopharmaceutical applications. Therefore, after successfully delivering the prototype device and completing this SBIR phase II, the vision is for the small business to make a commercially viable psoralen/UV-A-based inactivation device by partnering with vaccine manufacturers. This device can be marketed for making psoralen-inactivated vaccines against a broad range of diseases caused by viruses including influenza, poliovirus, hantavirus and rabies virus. Availability of such a high-throughput psoralen-inactivation device for manufacturing the GMP product with a basic instrument manual with operating instructions to regulate/adjust the device parameters and anticipated troubleshooting guidelines (in accordance with FDA guidelines) will significantly advance the psoralen-inactivated whole vaccine platform as an agile vaccine platform against emerging infectious threats. This pathogen inactivation device can also be marketed for rapid inactivation of contaminants during biopharmaceuticals production such as recombinant proteins and other therapeutic agents. It can also be marketed to academic and environmental scientists for inactivating high containment (BSL-3 and BSL-4 laboratories) pathogens without degrading their surface proteins and antigens before bringing them out of the BSL-3 or BSL-4 lab for antigen discovery and characterization. A basic instrument manual with operating instructions to regulate/adjust the device parameters and anticipated troubleshooting guidelines.
- Raviprakash K, Sun P, Raviv Y, Luke T, Martin N, and Kochel T, Dengue virus photo-inactivated in presence of 1,5-iodonaphthylazide (INA) or AMT, a psoralen compound (4'-aminomethyl-trioxsalen) is highly immunogenic in mice. Hum Vaccin Immunother, 2013. 9(11): p. 2336-41. hvi-9-2336.pdf (nih.gov).
- Maves RC, Ore RM, Porter KR, and Kochel TJ, Immunogenicity and protective efficacy of a psoralen-inactivated dengue-1 virus vaccine candidate in Aotus nancymaae monkeys. Vaccine, 2011. 29(15): p. 2691-6 https://doi.org/10.1016/j.vaccine.2011.01.077.
- Sundaram AK, Ewing D, Blevins M, Liang Z, Sink S, Lassan J, Raviprakash K, Defang G, Williams M, Porter KR, and Sanders JW, Comparison of purified psoralen-inactivated and formalin-inactivated dengue vaccines in mice and nonhuman primates. Vaccine, 2020. 38(17): p. 3313-3320. https://doi.org/10.1016/j.vaccine.2020.03.008.
- Sundaram AK, Ewing D, Liang Z, Jani V, Cheng Y, Sun P, Raviprakash K, Wu S-J, Petrovsky N, Defang G, Williams M, and Porter KR, Immunogenicity of Adjuvanted Psoralen-Inactivated SARS-CoV-2 Vaccines and SARS-CoV-2 Spike Protein DNA Vaccines in BALB/c Mice. Pathogens, 2021. 10(5): p. 626. • DOI: 10.3390/pathogens10050626.
KEYWORDS: Psoralen inactivation of pathogens, UV-A irradiation, Psoralen-inactivated pathogen, Whole virus inactivated vaccine, Emerging infectious diseases, Agile vaccine platform