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At least 739 records · Page 41

Hybrid Sorbents for 129 I Capture from Contaminated Groundwater

Radioiodine ( 129 I) poses a risk to the environment due to its long half-life, toxicity, and mobility. It is found at the U.S. Department of Energy Hanford Site due to legacy releases of nuclear wastes to the subsurface where 129 I is predominantly present as iodate (IO 3 – ). To date, a cost-effective and scalable cleanup technology for 129 I has not been identified, with hydraulic containment implemented as the remedial approach. Here, novel high-performing sorbents for 129 I remediation with the capacity to reduce 129 I concentrations to or below the US Environmental Protection Agency (EPA) drinking water standard and procedures to deploy them in an ex-situ pump and treat (P&T) system are introduced. Furthermore, this includes implementation of hybridized polyacrylonitrile (PAN) beads for ex-situ remediation of IO 3 – -contaminated groundwater for the first time. Iron (Fe) oxyhydroxide and bismuth (Bi) oxyhydroxide sorbents were deployed on silica substrates or encapsulated in porous PAN beads. In addition, Fe–, cerium (Ce)–, and Bi–oxyhydroxides were encapsulated with anion-exchange resins. The PAN–bismuth oxyhydroxide and PAN–ferrihydrite composites along with Fe- and Ce-based hybrid anion-exchange resins performed well in batch sorption experiments with distribution coefficients for IO 3 – of >1000 mL/g and rapid removal kinetics. Of the tested materials, the Ce-based hybrid anion-exchange resin was the most efficient for removal of IO 3 – from Hanford groundwater in a column system, with 50% breakthrough occurring at 324 pore volumes. The functional amine groups on the parent resin and amount of active sorbent in the resin can be customized to improve the iodine loading capacity. These results highlight the potential for IO 3 – remediation by hybrid sorbents and represent a benchmark for the implementation of commercially available materials to meet EPA standards for cleanup of 129 I in a large-scale P&T system.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Monitoring and Acquisition Real-time System (MARS)

MARS is a graphical user interface (GUI) written in MATLAB and Java, allowing the user to configure and control the Scalable Parallel Architecture for Real-Time Acquisition and Analysis (SPARTAA) data acquisition system. SPARTAA not only acquires data, but also allows for complex algorithms to be applied to the acquired data in real time. The MARS client allows the user to set up and configure all settings regarding the data channels attached to the system, as well as have complete control over starting and stopping data acquisition. It provides a unique "Test" programming environment, allowing the user to create tests consisting of a series of alarms, each of which contains any number of data channels. Each alarm is configured with a particular algorithm, determining the type of processing that will be applied on each data channel and tested against a defined threshold. Tests can be uploaded to SPARTAA, thereby teaching it how to process the data. The uniqueness of MARS is in its capability to be adaptable easily to many test configurations. MARS sends and receives protocols via TCP/IP, which allows for quick integration into almost any test environment. The use of MATLAB and Java as the programming languages allows for developers to integrate the software across multiple operating platforms.

Holland, Corbin↗

Validation of Loci-Stream for Autogenous Pressurization of Cryogenic Propellant Tank

Autogenous pressurization of cryogenic propellant tanks eliminates the need to have an additional pressurant tank on the space vehicle, which is highly advantageous due to reduced vehicle mass and design complexity. Autogenous pressurization therefore is one of the key technologies for deep space exploration and long-term space missions. The complex interaction of thermal gradients, turbulence and phase change near the interface make the problem a challenging one to model. Nodal analysis tools and reduced order models are unable to capture the necessary physics. 3-D CFD analyses are necessary to fully characterize autogenous pressurization. CFD analyses pose their own difficulties. The requisite CFD tool to tackle this problem need to be modular with the ability to incorporate various physics models, efficient, and computationally scalable for simulating flight size tanks. NASA MSFC's Loci-Stream CFD tool along with the VOF module is a great candidate to fit this mold. We demonstrate our modeling approach and validation of Loci-Stream for predicting autogenous pressurization of a flight scale propellant tank in order for the solver to serve as a reliable design and analysis tool for NASA's CFM application needs. Liquid hydrogen tank pressurization tests carried out at the MSFC test stand 300 facilities provide reliable validation data for this purpose. These tests were modeled using the Loci-Stream solver with a newly implemented two-phase sharp interface treatment. We show that our modeling approach and CFD solver predict the autogenous pressurization phenomena satisfactorily, and document challenging aspects of modeling this problem.

cryogenic fluid management↗

Validation of Loci-Stream for Autogenous Pressurization of Cryogenic Propellant Tank

Autogenous pressurization of cryogenic propellant tanks eliminates the need to have an additional pressurant tank on the space vehicle, which is highly advantageous due to reduced vehicle mass and design complexity. Autogenous pressurization therefore is one of the key technologies for deep space exploration and long-term space missions. The complex interaction of thermal gradients, turbulence and phase change near the interface make the problem a challenging one to model. Nodal analysis tools and reduced order models are unable to capture the necessary physics. 3-D CFD analyses are necessary to fully characterize autogenous pressurization. CFD analyses pose their own difficulties. The requisite CFD tool to tackle this problem need to be modular with the ability to incorporate various physics models, efficient, and computationally scalable for simulating flight size tanks. NASA MSFC's Loci-Stream CFD tool along with the VOF module is a great candidate to fit this mold. We demonstrate our modeling approach and validation of Loci-Stream for predicting autogenous pressurization of a flight scale propellant tank in order for the solver to serve as a reliable design and analysis tool for NASA's CFM application needs. Liquid hydrogen tank pressurization tests carried out at the MSFC test stand 300 facilities provide reliable validation data for this purpose. These tests were modeled using the Loci-Stream solver with a newly implemented two-phase sharp interface treatment. We show that our modeling approach and CFD solver predict the autogenous pressurization phenomena satisfactorily, and document challenging aspects of modeling this problem.

CFM↗

Validation of Two Phase Flow Modeling Techniques in Loci-Stream using Axial Jet Pressure Control and Autogenous Pressurization Experiment

Autogenous pressurization of cryogenic propellant tanks eliminates the need to have an additional pressurant tank on the space vehicle, which is highly advantageous due to reduced vehicle mass and design complexity. Autogenous pressurization therefore is one of the key technologies for deep space exploration and long-term space missions. The complex interaction of thermal gradients, turbulence and phase change near the interface make the problem a challenging one to model. Nodal analysis tools and reduced order models are unable to capture the necessary physics. 3-D CFD analyses are necessary to fully characterize autogenous pressurization. CFD analyses pose their own difficulties. The requisite CFD tool to tackle this problem need to be modular with the ability to incorporate various physics models, efficient, and computationally scalable for simulating flight size tanks. NASA MSFC's Loci-Stream CFD tool along with the VOF module is a great candidate to fit this mold. We demonstrate our modeling approach and validation of Loci-Stream for predicting autogenous pressurization of a flight scale propellant tank in order for the solver to serve as a reliable design and analysis tool for NASA's CFM application needs. Liquid hydrogen tank pressurization tests carried out at the MSFC test stand 300 facilities provide reliable validation data for this purpose. These tests were modeled using the Loci-Stream solver with a newly implemented two-phase sharp interface treatment. We show that our modeling approach and CFD solver predict the autogenous pressurization phenomena satisfactorily, and document challenging aspects of modeling this problem.

cryogenic fluid management↗

Adsorption-based direct air capture using hierarchical porous composites prepared via confined-space crystallization

Capturing CO₂ at trace concentration remains a critical challenge in sustainable carbon management via adsorption, as conventional adsorbents suffer from low CO₂ selectivity, poor moisture tolerance, and energy-intensive regeneration requirements. Here, we report a hierarchical Ba²⁺-exchanged silicoaluminophosphate (Ba²⁺-CSAPO-34) composite synthesized via confined-space crystallization within an activated carbon matrix. Comprehensive characterization revealed a confined nucleation mechanism and the successful incorporation of Ba²⁺ active sites within the SAPO-34 framework, achieved via a two-step liquid ion-exchange protocol. The core-shell architecture combines the selective CO₂ binding of Ba²⁺-functionalized SAPO-34 with the hydrophobic protection of the carbon shell. Fixed-bed adsorption tests demonstrated strong CO₂ binding (at 500-2500 ppm), no roll-up, and effective suppression of water affinity, while maintaining high selectivity even at 90% relative humidity. A phenomenological adsorption model, validated against dynamic breakthrough data, accurately predicted dynamic adsorption behavior under real-world operating conditions, enabling rational process design for direct air capture (DAC) and closed-loop life support systems. Furthermore, these results establish Ba²⁺-CSAPO-34 as a scalable, moisture-resistant adsorbent that addresses key limitations in trace CO₂ capture, advancing practical implementation of carbon removal technologies.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

The extent of multiallelic, co‐editing of LIGULELESS1 in highly polyploid sugarcane tunes leaf inclination angle and enables selection of the ideotype for biomass yield

Summary Sugarcane ( Saccharum spp. hybrid) is a prime feedstock for commercial production of biofuel and table sugar. Optimizing canopy architecture for improved light capture has great potential for elevating biomass yield. LIGULELESS1 ( LG1 ) is involved in leaf ligule and auricle development in grasses. Here, we report CRISPR/Cas9‐mediated co‐mutagenesis of up to 40 copies/alleles of the putative LG1 in highly polyploid sugarcane (2 n = 100–120, x = 10–12). Next generation sequencing revealed co‐editing frequencies of 7.4%–100% of the LG1 reads in 16 of the 78 transgenic lines. LG1 mutations resulted in a tuneable leaf angle phenotype that became more upright as co‐editing frequency increased. Three lines with loss of function frequencies of ~12%, ~53% and ~95% of lg1 were selected following a randomized greenhouse trial and grown in replicated, multi‐row field plots. The co‐edited LG1 mutations were stably maintained in vegetative progenies and the extent of co‐editing remained constant in field tested lines L26 and L35. Next generation sequencing confirmed the absence of potential off targets. The leaf inclination angle corresponded to light transmission into the canopy and tiller number. Line L35 displaying loss of function in ~12% of the lg1 NGS reads exhibited an 18% increase in dry biomass yield supported by a 56% decrease in leaf inclination angle, a 31% increase in tiller number, and a 25% increase in internode number. The scalable co‐editing of LG1 in highly polyploid sugarcane allows fine‐tuning of leaf inclination angle, enabling the selection of the ideotype for biomass yield.

59 BASIC BIOLOGICAL SCIENCES↗

Tamper-Indicating Enclosures with Visually Obvious Tamper Response (Final Project Report)

Sandia National Laboratories is developing a new method for detecting penetration of tamper - indicating enclosures (TIEs). This method incorporates the use of "bleeding" materials (analogous to visually obvious, colorful bruised skin that doesn't heal) into the design of TIEs. As designed, it will allow inspectors to use simple visual observation to detect attempts to penetrate the external surfaces of a TIE, without providing adversaries the ability to repair damage. A material of this type can enhance tamper indication of current TIEs used to support treaty verification regimes. Current TIE inspections are time - consuming and rely on subjective visual assessment by an inspector, equipment such as eddy current or camera devices, or involve approaches that may be limited due to application environment. The complexities and requirements that volumetric sealing methods (or TIEs) must address are: (1) enclosures that are non - standard in size/shape; (2) enclosures that may be inspectorate - or facility - owned; (3) finding tamper attempts that are difficult and time consuming for an inspector to locate; (4) enclosures that are reliable and durable enough to survive the conditions that exist in the operating environment (including facility handling); and (5) methods that prevent adversaries from repairing penetrations. Early project R&D [1] focused on encapsulated transition metals. Due to the challenges associated with the transition metal - based approach, a mitigation approach was investigated resulting in two separate research paths — one that involves fabricating custom TIE molds that meet the specific (size and shape) needs of safeguards equipment a nd one that can be deployed as a sprayed on or painted coating to an existing TIE or surface. The "custom mold" approach is based on creating thin layers of materials that , when penetrated, expose an inner material to O 2 which causes an irreversible color change. The "in-situ coating" approach is based on applying a sensor solution containing color changing microcapsules that bleed when the microcapsule is ruptured. The anticipated benefits of this work are passive, flexible, scalable, robust , cost-effective TIEs with visually obvious responses to tamper attempts. This provides more efficient and effective monitoring , as inspectors will require little or no additional equipment and will be able to detect tamper without extensive time - consuming visual examination. Applications include custom TIEs (cabinets , equipment enclosures or seal bodies ), or spray-coating/painting onto facility-owned items, walls or structures, or circuit boards. The paper describes research and testing completed to-date on the method and integration of select system components.

36 MATERIALS SCIENCE↗

Transformative Efficiency and Automation in Modular Homes (TEAMH)

This report documents the Transformative Efficiency and Automation in Modular Homes (TEAMH) project, which evaluates the integration of advanced building envelope technologies and automation-assisted modular construction to improve residential energy performance and construction efficiency. The study investigates high-performance insulation systems, including vacuum insulation panels (VIPs), combined with light gauge steel (LGS) modular construction and factory automation. Laboratory testing, whole-building energy modeling across multiple climate zones, and factory demonstrations were conducted to assess thermal performance, energy savings, and production efficiency. Results indicate that upgraded envelope assemblies can achieve up to ~50% heating and ~34% cooling energy savings relative to IECC 2018 code-compliant homes, while automation-assisted construction can reduce wall assembly time by 24%–46% compared to conventional wood framing. The findings demonstrate the potential for scalable, high-performance modular homes that deliver significant energy savings with competitive projected costs.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Revamping Spacecraft Operational Intelligence

The EPOXI flight mission has been testing a new commercial system, Splunk, which employs data mining techniques to organize and present spacecraft telemetry data in a high-level manner. By abstracting away data-source specific details, Splunk unifies arbitrary data formats into one uniform system. This not only reduces the time and effort for retrieving relevant data, but it also increases operational visibility by allowing a spacecraft team to correlate data across many different sources. Splunk's scalable architecture coupled with its graphing modules also provide a solid toolset for generating data visualizations and building real-time applications such as browser-based telemetry displays.

Hwang, Victor↗

Initial Development of Fusion Magnet Simulation Capabilities for Performance and Safety Evaluation Using the MOOSE Framework

Fusion energy holds the promise of being a transformative technology as a carbon-neutral, sustainable source of energy. Whole device modeling and the development of fusion digital twins will be increasingly important for emerging fusion device concepts at both national laboratories and within the commercial fusion industry. However, meeting the challenge of whole device modeling of fusion energy devices requires robust, multiphysics, multiscale modeling and simulation technologies capable of running on large-scale supercomputers. Detailed analysis of individual systems at-scale is also required to ensure safe and efficient operation as well as provide the safety basis for future device designs and licensing activities. In a tokamak, toroidal and poloidal magnets confine and shape the fusion plasma to promote the fusion reaction. High plasma temperatures and high magnetic field requirements in modern design concepts (leading to high amounts of energy stored within each magnet) impose electrical, thermal, and mechanical loads on the magnet components, which in turn impacts the safety considerations of the magnet and their supporting systems. Idaho National Laboratory (INL) has a history of working in this space, including development and benchmarking of the Magnetic System Circuitry Analysis Program (MSCAP) and Magnet Arcing (MAGARC) codes to study magnet quench events; notably, MAGARC was used to study quenching during the ITER Engineering Design Activity. However, these legacy codes and capabilities are not parallel and scalable, and new tools are required for future advances in this area, which leads to the INL-developed Multiphysics Object-Oriented Simulation Environment (MOOSE) framework. Developed originally for fission reactor systems under United States Department of Energy, Office of Nuclear Energy modeling and simulation programs, the MOOSE framework has been well-suited to multiscale, multiphysics modeling and simulation needs for nuclear systems. The framework is open-source, well-tested, under continuous development and deployment, and developed to a Nuclear Quality Assurance, Level 1 software quality standard. MOOSE has also been used in the fusion space previously in several projects: INL’s Tritium Migration Analysis Program, Version 8 (TMAP8) for tritium migration, UK Atomic Energy Authority’s A Unified Resource for OpenMC (fusion) Reactor Applications (AURORA) code for fusion thermo-mechanical and neutronics analysis, and Argonne National Laboratory’s Cardinal for high-fidelity computational fluid dynamics and neutronics. However, to model superconducting magnets, several MOOSE enhancements are required: additions to the current MOOSE electromagnetic capabilities, new material libraries for superconductors of interest (such as YBCO), as well as fusion-specific models for thermo-mechanics. This talk will discuss initial development activities to build these capabilities in MOOSE, focusing on initial validation and benchmarking activities. Proposed coupling workflows and future work to support the simulation of fusion magnets and magnet structural assemblies for performance and safety evaluation in MOOSE will also be discussed.

70 - PLASMA PHYSICS AND FUSION TECHNOLOGY↗

An error-controlled adaptive time-stepping method for particle advancement in coupled CFD-DEM simulations

Coupled Computational-Fluid-Dynamics (CFD) and Discrete-Element-Method (DEM) models provide an accurate description of multiphase physical systems where a solid granular particle phase exists in an underlying gaseous continuous medium. The time integration of the granular phase in these simulations is typically handled using an explicit scheme with a constant time-step among all particles that is invariant in time to resolve inter-particle collisions. A locally third-order accurate adaptive time integration technique for particles that employs an embedded locally second-order scheme for error determination is presented in this work. The particle time-step size is dynamically adapted based on solution error, thus leading to significant savings in computational time. The efficacy of our scheme is quantified using four test cases of varying complexity (binary collision, homogeneous cooling system, fluidized bed and hopper discharge). The adaptive time-stepping method exhibits improved performance (~ 2–3 times in most of the cases studied) compared to three commonly used non-adaptive time-step methods (first-order Euler-explicit, second-order Adams-Bashforth and third-order Runge-Kutta schemes), while maintaining the same level of accuracy and parallel scalability.

42 ENGINEERING↗

Solar Arrays for Low-Irradiance Low-Temperature and High-Radiation Environments

This is the Base Period final report DRAFT for the JPL task 'Solar Arrays for Low-Irradiance Low-Temperature and High-Radiation Environments', under Task Plan 77-16518 TA # 21, for NASA's Extreme Environments Solar Power (EESP) project. This report covers the Base period of performance, 7/18/2016 through 5/2/2017.The goal of this project is to develop an ultra-high efficiency lightweight scalable solar array technology for low irradiance, low temperature and high-radiation (LILT/Rad) environments. The benefit this technology will bring to flight systems is a greater than 20 reduction in solar array surface area, and a six-fold reduction in solar array mass and volume. The EESP project objectives are summarized in the 'NRA Goal' column of Table 1. Throughout this report, low irradiance low temperature (LILT) refers to 5AU -125 C test conditions; beginning of life (BOL) refers to the cell state prior to radiation exposure; and end of life (EOL) refers to the test article condition after exposure to a radiation dose of 4e15 1MeV e(-)/cm(exp 2).

Boca, Andreea↗

TPS to Enable a Uranus Orbiter Probe

Recent mission studies point to delivery of the probe either at Uranus or at Neptune will require 3-D Woven TPS, either the two layer HEEET or the single layer 3MDCP. Recent studies point to 3MDCP being more mass efficient. The 3MDCP capability being developed to support MSR EES is sufficient if the probe size is 1.25 m or less. If the prove diameter is larger than 1.25 m, a possibility then Dual layer HEEET which is scalable is at TRL 6 is certified and ready. It is possible to develop a seam approach that would allow 3MDCP and the mass savings could warrant development of a seam capability. Depending on the outcome of the planned Flagship mission study that Ames plans to support, Ames is already working on a concept that can be developed and demonstrated in the coming years. If in case Aerocapture is demonstrated through a Earth Flight Test and becomes part of the mission design for Uranus, this makes delivery of the probes from orbit easier. Whether the probe is to delivered from Orbit or not, still 3-D Woven (HEEET or 3MDCP) is the only TPS qualified to enable the Ice Giant in situ probes.

Matthew J Gasch↗

Regression Verification Using Impact Summaries

Regression verification techniques are used to prove equivalence of syntactically similar programs. Checking equivalence of large programs, however, can be computationally expensive. Existing regression verification techniques rely on abstraction and decomposition techniques to reduce the computational effort of checking equivalence of the entire program. These techniques are sound but not complete. In this work, we propose a novel approach to improve scalability of regression verification by classifying the program behaviors generated during symbolic execution as either impacted or unimpacted. Our technique uses a combination of static analysis and symbolic execution to generate summaries of impacted program behaviors. The impact summaries are then checked for equivalence using an o-the-shelf decision procedure. We prove that our approach is both sound and complete for sequential programs, with respect to the depth bound of symbolic execution. Our evaluation on a set of sequential C artifacts shows that reducing the size of the summaries can help reduce the cost of software equivalence checking. Various reduction, abstraction, and compositional techniques have been developed to help scale software verification techniques to industrial-sized systems. Although such techniques have greatly increased the size and complexity of systems that can be checked, analysis of large software systems remains costly. Regression analysis techniques, e.g., regression testing [16], regression model checking [22], and regression verification [19], restrict the scope of the analysis by leveraging the differences between program versions. These techniques are based on the idea that if code is checked early in development, then subsequent versions can be checked against a prior (checked) version, leveraging the results of the previous analysis to reduce analysis cost of the current version. Regression verification addresses the problem of proving equivalence of closely related program versions [19]. These techniques compare two programs with a large degree of syntactic similarity to prove that portions of one program version are equivalent to the other. Regression verification can be used for guaranteeing backward compatibility, and for showing behavioral equivalence in programs with syntactic differences, e.g., when a program is refactored to improve its performance, maintainability, or readability. Existing regression verification techniques leverage similarities between program versions by using abstraction and decomposition techniques to improve scalability of the analysis [10, 12, 19]. The abstractions and decomposition in the these techniques, e.g., summaries of unchanged code [12] or semantically equivalent methods [19], compute an over-approximation of the program behaviors. The equivalence checking results of these techniques are sound but not complete-they may characterize programs as not functionally equivalent when, in fact, they are equivalent. In this work we describe a novel approach that leverages the impact of the differences between two programs for scaling regression verification. We partition program behaviors of each version into (a) behaviors impacted by the changes and (b) behaviors not impacted (unimpacted) by the changes. Only the impacted program behaviors are used during equivalence checking. We then prove that checking equivalence of the impacted program behaviors is equivalent to checking equivalence of all program behaviors for a given depth bound. In this work we use symbolic execution to generate the program behaviors and leverage control- and data-dependence information to facilitate the partitioning of program behaviors. The impacted program behaviors are termed as impact summaries. The dependence analyses that facilitate the generation of the impact summaries, we believe, could be used in conjunction with other abstraction and decomposition based approaches, [10, 12], as a complementary reduction technique. An evaluation of our regression verification technique shows that our approach is capable of leveraging similarities between program versions to reduce the size of the queries and the time required to check for logical equivalence. The main contributions of this work are: - A regression verification technique to generate impact summaries that can be checked for functional equivalence using an off-the-shelf decision procedure. - A proof that our approach is sound and complete with respect to the depth bound of symbolic execution. - An implementation of our technique using the LLVMcompiler infrastructure, the klee Symbolic Virtual Machine [4], and a variety of Satisfiability Modulo Theory (SMT) solvers, e.g., STP [7] and Z3 [6]. - An empirical evaluation on a set of C artifacts which shows that the use of impact summaries can reduce the cost of regression verification.

Backes, John↗

Proteome-wide analysis of protein stability in Escherichia coli under acid stress

Knowledge of protein acid sensitivity remains sparse and is largely derived from low-throughput, enzyme-specific assays. We used a scalable framework to map acid stability across the Escherichia coli proteome to assess the acid stability of 1,675 unique proteins, estimating pH 50 values for over 90% of them. The parameter pH50 was defined as the pH value at which only 50% of the initial protein remains in solution following acid treatment. Proteome-wide pH 50 values ranged from 2.28 to 6.33 (median 5.11). Approximately 9% of detected proteins remained stable across all tested pH conditions. Our results align with published data and the assay of citrate synthase (GltA) performed here. Protein acid stability differed significantly by subcellular localization: periplasmic proteins were relatively more abundant in the acid-stable group, cytoplasmic proteins were abundant at pH 50 values 4.5–5.5, and inner membrane proteins at higher pH 50 between 5.5 and 6.0. Outer membrane proteins were too few to draw strong conclusions regarding enrichment within specific pH 50 groups. Notably, the periplasmic binding protein of the molybdate ABC transporter (ModA), was enriched after incubation at low pH. Estimated pH 50 values showed no correlation with protein isoelectric point and molecular weight. Together, this work provides the first proteome-wide map of protein acid stability and establishes a general framework for studying different chemical stressors.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A cell-based Papain-like Protease (PLpro) activity assay for rapid detection of active SARS-CoV-2 infections and antivirals

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its variants are a continuous threat to human life. An urgent need remains for simple and fast tests that reliably detect active infections with SARS-CoV-2 and its variants in the early stage of infection. Here we introduce a simple and rapid activity-based diagnostic (ABDx) test that identifies SARS-CoV-2 infections by measuring the activity of a viral enzyme, Papain-Like protease (PLpro). The test system consists of a peptide that fluoresces when cleaved by SARS PLpro that is active in crude, unprocessed lysates from human tongue scrapes and saliva. Test results are obtained in 30 minutes or less using widely available fluorescence plate readers, or a battery-operated portable instrument for on-site testing. Proof-of-concept was obtained in a study on clinical specimens collected from patients with COVID-19 like symptoms who tested positive (n = 10) or negative (n = 10) with LIAT RT-PCR using nasal mid turbinate swabs. When saliva from these patients was tested with in-house endpoint RT-PCR, 17 were positive and only 5 specimens were negative, of which 2 became positive when tested 5 days later. PLpro activity correlated in 17 of these cases (3 out of 3 negatives and 14 out of 16 positives, with one invalid specimen). Despite the small number of samples, the agreement was significant (p value = 0.01). Two false negatives were detected, one from a sample with a late Ct value of 35 in diagnostic RT-PCR, indicating that an active infection was no longer present. The PLpro assay is easily scalable and expected to detect all viable SARS-CoV-2 variants, making it attractive as a screening and surveillance tool. Additionally, we show feasibility of the platform as a new homogeneous phenotypic assay for rapid screening of SARS-CoV-2 antiviral drugs and neutralizing antibodies.

60 APPLIED LIFE SCIENCES↗