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Identifying pharmacological probes for human proteins represents a key opportunity to accelerate the discovery of new therapeutics. High-content screening approaches to expand the ligandable proteome offer the potential to expedite the discovery of novel chemical probes to study protein function. Screening libraries of reactive fragments by chemoproteomics offers a compelling approach to ligand discovery, however, optimising sample throughput, proteomic depth, and data reproducibility remains a key challenge. We report a versatile, label-free quantification proteomics platform for competitive profiling of cysteine-reactive fragments against the native proteome. This high-throughput platform combines SP4 plate-based sample preparation with rapid chromatographic gradients. Data-independent acquisition performed on a Bruker timsTOF Pro 2 consistently identified ~23,000 cysteine sites per run, with a total of ~32,000 cysteine sites profiled in HEK293T and Jurkat lysate. Crucially, this depth in cysteinome coverage is met with high data completeness, enabling robust identification of liganded proteins. In this study, 80 reactive fragments were screened in two cell lines identifying >400 ligand-protein interactions. Hits were validated through concentration-response experiments and the platform was utilised for hit expansion and live cell experiments. This label-free platform represents a significant step forward in high-throughput proteomics to evaluate ligandability of cysteines across the human proteome. This paper presents a label-free chemoproteomics platform using data-independent acquisition to profile covalent fragment binding across the human proteome. The platform offers high reproducibility and data completeness, identifying >400 protein-ligand interactions for probe development.

Deubiquitinating enzymes (DUBs) are key regulators of cellular homoeostasis, and their dysregulation is associated with several human diseases. The ovarian tumour protease (OTU) family of DUBs are biochemically well-characterised and of therapeutic interest, yet only a few tool compounds exist to study their cellular function and therapeutic potential. Here we present a chemoproteomics fragment screening platform for identifying novel DUB-specific hit matter, that combines activity-based protein profiling with high-throughput chemistry direct-to-biology optimisation to enable rapid elaboration of initial fragment hits against OTU DUBs. Applying these approaches, we identify an enantioselective covalent fragment for OTUD7B, and validate it using chemoproteomics and biochemical DUB activity assays. The ovarian tumour protease (OTU) family of deubiquitinating enzymes (DUBs) are biochemically well-characterised and of therapeutic interest, however, only a few tool compounds exist to study their cellular function and therapeutic potential. Here, the authors present a chemoproteomics fragment screening platform for identifying DUB-specific hit matter, which combines activity-based protein profiling with high-throughput optimisation to enable rapid elaboration of initial fragment hits against OTU DUBs, and identify an enantioselective chloroacetamide-based covalent fragment for OTUD7B.

Identifying pharmacological probes for human proteins represents a key opportunity to accelerate the discovery of new therapeutics. High-content screening approaches to expand the ligandable proteome offer the potential to expedite the discovery of novel chemical probes to study protein function. Screening libraries of reactive fragments by chemoproteomics offers a compelling approach to ligand discovery, however, optimising sample throughput, proteomic depth, and data reproducibility remains a key challenge. We report a versatile, label-free quantification proteomics platform for competitive profiling of cysteine-reactive fragments against the native proteome. This high-throughput platform combines SP4 plate-based sample preparation with rapid chromatographic gradients. Data-independent acquisition performed on a Bruker timsTOF Pro 2 consistently identified ∼23,000 cysteine sites per run, with a total of ∼32,000 cysteine sites profiled in HEK293T and Jurkat lysate. Crucially, this depth in cysteinome coverage is met with high data completeness, enabling robust identification of liganded proteins. In this study, 80 reactive fragments were screened in two cell lines identifying >400 ligand-protein interactions. Hits were validated through concentration-response experiments and the platform was utilised for hit expansion and live cell experiments. This label-free platform represents a significant step forward in high-throughput proteomics to evaluate ligandability of cysteines across the human proteome.

Chemoproteomics is a powerful method capable of detecting interactions between small molecules and the proteome, however its use as a high-throughput screening method for chemical libraries has so far been limited. To address this need, we have further developed a chemoproteomics workflow to screen cysteine reactive covalent fragments in cell lysates against the deubiquitinating (DUB) enzymes using activity-based protein profiling. By using targeted ubiquitin probes, we have addressed sensitivity and affinity limitations, enabling target identification and covalent fragment library profiling in a 96-well plate format. The use of data independent acquisition (DIA) methods for MS analysis combined with automated Evosep liquid chromatography (LC) reduced instrument runtimes to 21 minutes per sample and simplified the workflow. In this proof-of-concept study, we have profiled 138 covalent fragments against 57 DUB proteins and validated four hit fragments against OTUD7B and UCHL3 through site identification experiments and orthogonal biochemical activity assays.Competing Interest StatementThe authors have declared no competing interest.

Recently, the appeal of Hybrid Energy Storage Systems (HESSs) has been growing in multiple application fields, such as charging stations, grid services, and microgrids. HESSs consist of an integration of two or more single Energy Storage Systems (ESSs) to combine the benefits of each ESS and improve the overall system performance, e.g., efficiency and lifespan. Most recent studies on HESS mainly focus on power management and coupling between the different ESSs without a particular interest in a specific type of ESS. Over the last decades, Redox-Flow Batteries (RFBs) have received significant attention due to their attractive features, especially for stationary storage applications, and hybridization can improve certain characteristics with respect to short-term duration and peak power availability. Presented in this paper is a comprehensive overview of the main concepts of HESSs based on RFBs. Starting with a brief description and a specification of the Key Performance Indicators (KPIs) of common electrochemical storage technologies suitable for hybridization with RFBs, HESS are classified based on battery-oriented and application-oriented KPIs. Furthermore, an optimal coupling architecture of HESS comprising the combination of an RFB and a Supercapacitor (SC) is proposed and evaluated via numerical simulation. Finally, an in-depth study of Energy Management Systems (EMS) is conducted. The general structure of an EMS as well as possible application scenarios are provided to identify commonly used control and optimization parameters. Therefore, the differentiation in system-oriented and application-oriented parameters is applied to literature data. Afterwards, state-of-the-art EMS optimization techniques are discussed. As an optimal EMS is characterized by the prediction of the system’s future behavior and the use of the suitable control technique, a detailed analysis of the previous implemented EMS prediction algorithms and control techniques is carried out. The study summarizes the key aspects and challenges of the electrical hybridization of RFBs and thus gives future perspectives on newly needed optimization and control algorithms for management systems.

The dual-ion “Saltwater Battery” based on aqueous electrolyte containing sodium ions and lithium ions is believed to be one of the safest and environmentally friendliest battery technologies. The anode consists of sodium titanium phosphate, whereas the cathode is spinel lithium manganese oxide. It has been reported that both materials can intercalate sodium and lithium ions depending on their availability in the electrolyte. This study aims to identify the dominant active species in “Saltwater Batteries” with dual-ion electrolyte. Therefore, cyclic voltammetry of single electrodes as well as full cell measurements are performed with electrolyte containing lithium sulfate, sodium sulfate or a mixture. Moreover, the study is complemented by ion analysis of the electrolyte, as well as X-ray diffraction of fresh and cycled electrodes at different states of charge. The results show that the cathode only (de)intercalates lithium ions, revealing that Li is the dominant active cathode species. The anode can react with both ions and undergoes a formation reaction accompanied by partial dissolution of sodium titanium phosphate. Nevertheless, cyclic voltammetry and full cell measurements indicate that lithium is also the dominant active species on the anode side. In conclusion, the dual-ion battery is dominated by lithium and shows a superior performance when removing sodium from electrolyte.

Respiratory Syncytial Virus (RSV) is a common cause of severe respiratory tract disease in infants, the elderly and immunocompromised patients. However, there is uncertainty as to how sample handling practices affect performance of tests to detect RSV. The aim of this study was to determine whether RSV RNA remains reliably detectable in nasopharyngeal/oropharyngeal (NP/OP) samples, saliva, and sputum samples over time. Respiratory samples were collected as part of a prospective observational study of acute lower respiratory tract disease (aLRTD) hospitalisations in adults in Bristol (UK). Samples that tested positive by PCR on receipt (0 h), were re-tested at 24 h having been stored at room temperature. We found that all but one of the samples PCR-positive for RSV at 0 h remained positive at 24 h, across all sample types and RSV strains. Ct values for NP/OP and saliva samples were significantly lower at 24 h than at 0 h, suggesting potential low-level viral replication in the samples. These results suggest that RSV tests can provide consistent results after a delay of up to 24 h following sample collection.

Background The burden of Respiratory Syncytial Virus (RSV) infection in adults is of interest in the context of recently-licensed vaccines. However, burden estimates are affected by test error associated with the testing platform, and number and type of samples tested. Methods We conducted a prospective cohort study of adults with acute lower respiratory tract disease (aLRTD) hospitalised in Bristol, UK, from April 2022–March 2023. RSV was detected by RT-PCR both by routine standard-of-care (SOC) testing, and by testing of additional nasopharyngeal swabs, saliva and sputum samples from a patient subset. Latent class analysis was used to quantify and adjust for test error rates, including effects of multiple testing. RSV test-positivity rates are reported, and after adjustment for test error, are used to calculate adult population incidence/1000 person-years. Results 6906/11445 aLRTD cases (60%) were tested and 251 were positive (3.6%; 251/6906). Test-positivity peaked in December (95%CI 7.9–12.7%). Among cases, 43% had pneumonia, 55% had non-pneumonic infection, 59% chronic respiratory disease exacerbations, and 16% heart failure. Test-positivity was highest in 75–84-year-olds, and 30-day mortality was highest in ≥ 75-year-olds (7.1%; 9/127). Due to low positivity-rates and imperfect specificity (0.98–1.00), test-positivity (3.6%) overestimated inferred true prevalence (2.3%). After adjustment for test error, we estimate overall adult population incidence/1000-person-years to be 0.33 (0.21–0.49), and 2.02 (1.10–3.06) in ≥ 75-year-olds. Conclusions RSV contributes significantly to hospitalised adult aLRTD, particularly among the elderly. The implementation of effective RSV vaccines could reduce morbidity, mortality and associated costs of disease. Adult RSV burden accuracy is improved by adjustment for test characteristics due to the impact of imperfect specificity when positivity-rates are low, and this is particularly important for out-of-season estimates. Multiple samples can improve burden estimation accuracy only when tests have near-perfect specificity.