When will the immune-stimulating antibody conjugates (ISACs) be transferred from bench to bedside?

Immunostimulatory antibody conjugates (ISACs) as a promising new generation of targeted therapeutic antibody-drug conjugates (ADCs), that not only activate innate immunity but also stimulate adaptive immunity, providing a dual therapeutic effect to eliminate tumor cells. However, several ISACs are still in the early stages of clinical development or have already failed. Therefore, it is crucial to design ISACs more effectively to overcome their limitations, including high toxicity, strong immunogenicity, long development time, and poor pharmacokinetics. This review aims to summarize the composition and function of ISACs, incorporating current design considerations and ongoing clinical trials. Additionally, the review delves into the current issues with ISACs and potential solutions, such as adjusting the drug-antibody ratio (DAR) to improve the bioavailability of ISACs. By leveraging the affinity and bioavailability-enhancing properties of bispecific antibodies, the utility between antibodies and immunostimulatory agents can be balanced. Commonly used immunostimulatory agents may induce systemic immune reactions, and BTK (Bruton's tyrosine kinase) inhibitors can regulate immunogenicity. Finally, the concept of grafting ADC's therapeutic principles is simple, but the combination of payload, linker, and targeted functional molecules is not a simple permutation and combination problem. The development of conjugate drugs faces more complex pharmacological and toxicological issues. Standing on the shoulders of ADC, the development and application scenarios of ISAC are endowed with broader space.

Multistatic Integrated Sensing and Communication System Based on Macro-Micro Cooperation.

A novel multistatic integrated sensing and communication (ISAC) system based on macro-micro cooperation for the sixth-generation (6G) mobile network is proposed. Instead of using macrosites at both the transmitter and receiver sides, microsites are considered as receivers in cooperative sensing. This system is important since microsites can be deployed more flexibly to reduce their distances to the sensing objects, providing better coverage for sensing service. In this work, we first analyze the deployment problem of microsites, which can be deployed along the radius and azimuth angle to cover macrosite cells. The coverage area of each microsite is derived in terms of its position in the cell. Then, we describe an efficient estimating approach for obtaining the position and velocity of sensing objects in the macrosite cell. By choosing multiple microsites around the targeted sensing area, joint data processing with an efficient optimization method is also provided. Simulation results show that the multistatic ISAC system employing macro-micro cooperation can improve the position and velocity estimation accuracy of objects compared to systems employing macrosite cooperation alone, demonstrating the effectiveness and potential for implementing the proposed system in the 6G mobile network.

Joint Active and Passive Beamforming in RIS-Assisted Secure ISAC Systems.

This paper investigates joint beamforming in a secure integrated sensing and communications (ISAC) system assisted by reconfigurable intelligent surfaces (RIS). The system communicates with legitimate downlink users, detecting a potential target, which is a potential eavesdropper attempting to intercept the downlink communication information from the base station (BS) to legitimate users. To enhance the physical-layer secrecy of the system, we design and introduce interference signals at the BS to disrupt eavesdroppers' attempts to intercept legitimate communication information. The BS simultaneously transmits communication and interference signals, both utilized for communication and sensing to guarantee the sensing and communication quality. By jointly optimizing the BS active beamformer and the RIS passive beamforming matrix, we aim to maximize the achievable secrecy rate and radiation power of the system. We develop an effective scheme to find the active beamforming matrix through fractional programming (FP) and semi-definite programming (SDP) techniques and obtain the RIS phase shift matrix via a local search technique. Simulation results validate the effectiveness of the proposed methods in enhancing communication and sensing performance. Additionally, the results demonstrate the effectiveness of introducing the interference signals and RIS in enhancing the physical-layer secrecy of the ISAC system.

Integrated Communication, Sensing, and Computation Framework for 6G Networks.

In the sixth generation (6G) era, intelligent machine network (IMN) applications, such as intelligent transportation, require collaborative machines with communication, sensing, and computation (CSC) capabilities. This article proposes an integrated communication, sensing, and computation (ICSAC) framework for 6G to achieve the reciprocity among CSC functions to enhance the reliability and latency of communication, accuracy and timeliness of sensing information acquisition, and privacy and security of computing to realize the IMN applications. Specifically, the sensing and communication functions can merge into unified platforms using the same transmit signals, and the acquired real-time sensing information can be exploited as prior information for intelligent algorithms to enhance the performance of communication networks. This is called the computing-empowered integrated sensing and communications (ISAC) reciprocity. Such reciprocity can further improve the performance of distributed computation with the assistance of networked sensing capability, which is named the sensing-empowered integrated communications and computation (ICAC) reciprocity. The above ISAC and ICAC reciprocities can enhance each other iteratively and finally lead to the ICSAC reciprocity. To achieve these reciprocities, we explore the potential enabling technologies for the ICSAC framework. Finally, we present the evaluation results of crucial enabling technologies to show the feasibility of the ICSAC framework.

Selected Technical Aspects of Molecular Allergy Diagnostics.

Diagnosis of allergic diseases is a complex, multi-stage process. It often requires the use of various diagnostic tools. The in vitro diagnostics (IVD), which includes various laboratory tests, is one of the stages of this process. Standard laboratory tests include the measurement of the serum concentration of specific immunoglobulin E (sIgE) for selected allergens, full allergen extracts and/or single allergen components (molecules). The measurement of IgE sIgE to the allergen components is called molecular allergy diagnosis. During the standard laboratory diagnostic process, various models of immunochemical tests are used, which enable the measurement of sIgE for single allergens (one-parameter tests, singleplex) or IgE specific for many different allergens (multi-parameter tests, multiplex) in one test. Currently, there are many different test kits available, validated for IVD, which differ in the method type and allergen profile. The aim of the manuscript is to present various technical aspects related to modern allergy diagnostics, especially in the area of molecular allergy diagnostics.

Generation and structure-activity relationships of novel imidazo-thienopyridine based TLR7 agonists: application as payloads for immunostimulatory antibody drug-conjugates.

Pairing immunostimulatory small molecules with the targeting capability of an antibody has emerged as a novel therapeutic modality with the potential to treat a variety of solid tumors. A series of compounds based on an imidazo-thienopyridine scaffold were synthesized and tested for their ability to agonize the innate immune sensors toll-like receptor 7 and 8 (TLR7/8). Structure-activity relationship (SAR) studies revealed that certain simple amino-substituents could enable TLR7 agonism at low nanomolar concentrations. Drug-linkers containing either payload 1 or 20h were conjugated to the HER2-targeting antibody trastuzumab at the interchain disulfide cysteine residues using a cleavable valine-citrulline dipeptide linker and stochastic thiol-maleimide chemistry. In vitro, these immune-stimulating antibody drug-conjugates (ADCs) were found to induce cytokine release in a murine splenocyte assay when co-cultured with the HER2-high NCI-N87 cancer cell line. In vivo, tumor regression was observed with a single dose in an NCI-N87 gastric carcinoma xenograft model in BALB/c nude mice.

Would the choice of multiplex platform impact the management of the allergic patient? A first approach focusing on LTPs.

BACKGROUND: In the Mediterranean area, patients with LTP syndrome who are sensitized to multiple allergens are often tested for sIgE using multiplex platforms. The results obtained from different commercial platforms are not interchangeable, so it is important to compare and validate the platform selected for use. The objective of this study is to compare and validate the performance of the ImmunoCAP ISAC E112i and the macroarray ALEX2 in our daily practice. METHODS: From August 2021 to March 2022, we tested 20 random serum samples from polysensitized patients using the ALEX2 test (MADx) and ImmunoCAP tIgE and ISAC E112i (Thermo Fisher Scientific). We compared the total IgE (tIgE) and sIgE levels for shared allergens. RESULTS: The heatmap generally showed more intense results for ISAC. The overall correlation was good, but some exceptions were noted. The main discrepancies were found for Ole e 7, which was positive for 11 patients in ISAC but negative for all patients in ALEX2, and for nut LTPs, for which ISAC showed a threefold higher detection rate for Ara h 9 and a fivefold higher detection rate for Cor a 8 and Jug r 3 compared to ALEX2. The regression model showed no interchangeability of tIgE results. CONCLUSIONS: Despite our small sample size and the complexity of comparing a quantitative and a semi-quantitative platform, our results suggest that patient diagnosis and management can be influenced by the platform used. Therefore, our findings must be taken into consideration when choosing a platform to use for some profiles of LTP-polysensitized patients, even though more data is needed.

In-City Rain Mapping from Commercial Microwave Links-Challenges and Opportunities.

Obtaining accurate rainfall measurements is highly important in urban areas, having a significant impact on different aspects in city life. Opportunistic rainfall sensing utilizing measurements collected by existing microwave and mmWave-based wireless networks has been researched in the last two decades and can be considered as an opportunistic integrated sensing and communication (ISAC) approach. In this paper, we compare two methods that utilize received signal level (RSL) measurements obtained by an existing smart-city wireless network deployed in the city of Rehovot, Israel, for rain estimation. The first method is a model-based approach using the RSL measurements from short links, in which two design parameters are calibrated empirically. This method is combined with a known wet/dry classification method, which is based on the rolling standard deviation of the RSL. The second method is a data-driven approach, based on a recurrent neural network (RNN), which is trained to estimate rainfall and classify wet/dry periods. We compare the results of rainfall classification and estimation from both methods and show that the data-driven approach slightly outperforms the empirical model and that the improvement is most significant for light rainfall events. Furthermore, we apply both methods to construct high-resolution 2D maps of accumulated rainfall in the city of Rehovot. The ground-level rainfall maps constructed over the city area are compared for the first time with weather radar rainfall maps obtained from the Israeli Meteorological Service (IMS). The rain maps generated by the smart-city network are found to be in agreement with the average rainfall depth obtained from the radar, demonstrating the potential of using existing smart-city networks as a source for constructing 2D high-resolution rainfall maps.

FDSS-Based DFT-s-OFDM for 6G Wireless Sensing.

Integrated sensing and communications (ISAC) is emerging as a key technology of 6G. Owing to the low peak-to-average power ratio (PAPR) property, discrete Fourier transform spread orthogonal frequency-division multiplexing (DFT-s-OFDM) is helpful to improve the sensing range and suitable for high-frequency transmission. However, compared to orthogonal frequency-division multiplexing (OFDM), the sensing accuracy of DFT-s-OFDM is relatively poor. In this paper, frequency-domain spectral shaping (FDSS) is adopted to enhance the performances of DFT-s-OFDM including sensing accuracy and PAPR by adjusting the correlation of signals. Specifically, we first establish a signal model for the ISAC system, followed by the description of performance indicators. Then, we analyze the influence of amplitude fluctuation of frequency domain signals on sensing performance, which shows the design idea of FDSS-enhanced DFT-s-OFDM. Further, a FDSS-enhanced DFT-s-OFDM framework is introduced for ISAC, where two types of FDSS filters including a pre-equalization filter and an isotropic orthogonal transform algorithm (IOTA) filter are designed. The simulation results show that the proposed scheme can obtain about 4 dB performance gain in terms of sensing accuracy over DFT-s-OFDM. In addition, FDSS-enhanced DFT-s-OFDM can significantly reduce PAPR and improve the power amplifier efficiency.

Doubly Constrained Waveform Optimization for Integrated Sensing and Communications.

This paper investigates threshold-constrained joint waveform optimization for an integrated sensing and communication (ISAC) system. Unlike existing studies, we employ mutual information (MI) and sum rate (SR) as sensing and communication metrics, respectively, and optimize the waveform under constraints to both metrics simultaneously. This provides significant flexibility in meeting system performance. We formulate three different optimization problems that constrain the radar performance only, the communication performance only, and the ISAC performance, respectively. New techniques are developed to solve the original problems, which are NP-hard and cannot be directly solved by conventional semi-definite programming (SDP) techniques. Novel gradient descent methods are developed to solve the first two problems. For the third non-convex optimization problem, we transform it into a convex problem and solve it via convex toolboxes. We also disclose the connections between three optimizations using numerical results. Finally, simulation results are provided and validate the proposed optimization solutions.

Joint Method of Moments (JMoM) and Successive Moment Cancellation (SMC) Multiuser Time Synchronization for ZP-OFDM-Based Waveforms Applicable to Joint Communication and Sensing.

It has been recently shown that zero padding (ZP)-orthogonal frequency-division multiplexing (OFDM) is a promising candidate for 6G wireless systems requiring joint communication and sensing. In this paper, we consider a multiuser uplink scenario where users are separated in power domain, i.e., non-orthogonal multiple access (NOMA), and use ZP-OFDM signals. The uplink transmission is grant-free and users are allowed to transmit asynchronously. In this setup, we address the problem of time synchronization by estimating the timing offset (TO) of all the users. We propose two non-data-aided (NDA) estimators, i.e., the joint method of moment (JMoM) and the successive moment cancellation (SMC), that employ the periodicity of the second order moment (SoM) of the received samples for TO estimation. Moreover, the coding assisted (CA) version of the proposed estimators, i.e., CA-JMoM and CA-SMC, are developed for the case of short observation samples. We also extend the proposed estimators to multiuser multiple-input multiple-output (MIMO) systems. The effectiveness of the proposed estimators is evaluated in terms of lock-in probability under various practical scenarios. Simulation results show that the JMoM estimator can reach the lock-in probability of one for the moderate range of Eb/N0 values. While existing NDA TO estimators in the literature either offer low lock-in probability, high computational complexity that prevents them from being employed in MIMO systems, or are designed for single-user scenarios, the proposed estimators in this paper address all of these issues.

A Novel Joint Channel Estimation and Symbol Detection Receiver for Orthogonal Time Frequency Space in Vehicular Networks.

A vehicular network embodies a specialized variant of wireless network systems, characterized by its capability to facilitate inter-vehicular communication and connectivity with the encompassing infrastructure. With the rapid development of wireless communication technology, high-speed and reliable communication has become increasingly important in vehicular networks. It has been demonstrated that orthogonal time frequency space (OTFS) modulation proves effective in addressing the challenges posed by high-mobility environments, as it transforms the time-varying channels into the delay-Doppler domain. Motivated by this, in this paper, we focus on the theme of integrated sensing and communication (ISAC)-assisted OTFS receiver design, which aims to perform sensing channel estimation and communication symbol detection. Specifically, the estimation of the sensing channel is accomplished through the utilization of a deep residual denoising network (DRDN), while the communication symbol detection is performed by orthogonal approximate message passing (OAMP) processing. The numerical results demonstrate that the proposed ISAC system exhibits superior performance and robustness compared to traditional methods, with a lower complexity as well. The proposed system has great potential for future applications in wireless communication systems, especially in challenging scenarios with high mobility and interference.

Microarray Analysis Confirms ImmunoCAP-Fluorescence Enzyme Immunoassay Results on Specific IgE in Patients with Atopic Dermatitis and Suspected Birch Pollen-Related Food Allergy.

BACKGROUND: Previous studies demonstrated that birch pollen-related foods can cause late eczematous responses in birch pollen-sensitized patients with atopic dermatitis (AD). However, suitable markers to predict birch pollen-related food allergy in patients with AD are still lacking. OBJECTIVE: We evaluated the correlation of the results from ImmunoCAP® fluorescence enzyme immunoassay (FEIA) singleplex and ImmunoCAP® immuno solid-phase allergen chip (ISAC) multiplex system in AD patients and investigated the diagnostic validity of allergen microarray analysis, measuring specific IgE (sIgE) with ImmunoCAP® ISAC to predict birch pollen-related food allergy in patients with AD. METHODS: A total of 19 children and adults with AD, existing IgE-mediated birch pollen sensitization, and suspected birch pollen-related food allergy underwent a double-blind placebo-controlled food challenge (DBPCFC) in the clinical routine. Total and sIgE levels to birch pollen, Bet v 1, Bet v 2, and birch pollen-related foods (apple, carrot, celery, and hazelnut) were determined prior to the DBPCFC by ImmunoCAP®-FEIA. Additionally, allergen microarray ImmunoCAP® ISAC analysis was performed. Data were analyzed retrospectively. RESULTS: Twelve out of 19 patients (63% responders) experienced an allergic reaction upon DBPCFC. Overall, 7 patients (37%) developed a significant deterioration of AD with a median increase of 12.4 points in the scoring of atopic dermatitis (SCORAD) index (range 10.0-15.7). Oral allergy syndrome was the predominant immediate-type symptom (n = 11/12 responders). There were no differences in sensitization frequencies regarding allergens of the pathogenesis-related protein family 10 between responders and non-responders. In all patients, correlation of IgE levels determined with ImmunoCAP® ISAC and ImmunoCAP®-FEIA, respectively, was significant with high correlation coefficients regarding birch pollen allergen extract, rBet v 1, and rBet v 2 (rs > 0.8, p < 0.001) and lower but also significant correlation coefficients regarding food allergens (rs < 0.8, p < 0.05-<0.001). CONCLUSION: ImmunoCAP® ISAC microarray allows displaying a differentiated sensitization profile in birch pollen-sensitized patients with AD. However, IgE-mediated sensitization against birch pollen-related allergens revealed by the allergen multiplex system does not predict late eczematous reactions upon DBPCFC with birch pollen-related foods.

A comprehensive comparison between ISAC and ALEX2 multiplex test systems.

OBJECTIVES: Diagnosis of type I hypersensitivity is based on anamnesis, provocation as well as blood- and skin testing. Multiplex specific IgE (sIgE) testing enables determination of sIgE antibodies against multiple recombinant or purified natural allergen components. The aim of this study was to evaluate the performance of the novel ALEX2® (Allergy Explorer, ALEX2 test introduced on the market November 2019) multiplex platform and to compare it with the ImmunoCAP ISAC® test system. METHODS: Serum samples of 49 patients, routinely determined with ISAC, were selected based on positive results covering in total most of the 112 ISAC components. Cohen's kappa, negative percent agreement (NPA), and positive percent agreement (PPA) of ALEX2 data compared to ISAC data (as a non-reference standard) were computed for those allergen components present on both platforms (n=103). Furthermore, in some samples sIgE results against allergen extracts and/or -components tested with either ImmunoCAP® (ThermoFisher) or IMMULITE® (Siemens) were available and compared to ALEX2 results. RESULTS: The overall agreement between ISAC and ALEX2 common allergen components was 94%. NPA and PPA were respectively 95 and 90%. Kappa values differed for specific allergen groups and varied between 0.60 and 0.92 showing moderate to almost perfect agreement. Of the qualitative discrepancies between ALEX2 and ISAC, 59% were related to weak positive results i.e. results under 1 kUA/L or 1 ISU, respectively. CONCLUSIONS: The method comparison between ISAC and ALEX2 multiplex tests showed a high concordance for those allergen components present on both platforms.

ALEX versus ISAC multiplex array in analyzing food allergy in atopic children.

ALEX multiplex array is a relatively new multiplex allergy test which analyses more than 120 allergen extracts and 170 molecular components. ISAC is the most used and studied multiplex array to date, offering 112 molecular components. In ten atopic children with multiple food allergies good agreement was observed between ALEX and ISAC sIgE results for nearly all shared food components. Presence of larger number of allergens in ALEX could help clinicians to improve personalized dietary advice. However more positive sensitizations with unknown clinical relevance were found by ALEX, potentially increasing clinical complexity. Pediatric allergists should be aware of this, especially in young atopic children with (severe) eczema who have not introduced all sorts of food yet.

On SFBC Schemes for Enabling Virtual Array Concept in Monostatic ISAC Scenarios.

The integrated sensing and communication (ISAC) paradigm is being proposed for 6G as a new feature of the physical layer (PHY), for tackling dual-functional applications, i.e., demanding radio-sensing and communication functions, such as the Internet of Things (IoT) and autonomous driving systems. This work considers the integration of sensing and communications functionalities in a unique platform. To achieve this goal, the use of orthogonal space frequency block codes (SFBC) is proposed. SFBC code orthogonality enables both the separation of communications data streams at a user terminal and the estimation of target parameters. The SFBC enhances the communications link diversity without requiring channel state information knowledge at the transmitter and enable the virtual antenna array concept for enhancing the direction-finding resolution. The use of different SFBCs provides a tradeoff between achieved diversity and sensing resolution. For example, an Alamouti code, applicable for the case with two transmitting antennas, duplicates sensing resolution and achieves a diversity order of two while the use of a Tarokh code, applicable for a scenario with four transmitting antennas, provides a fourfold better resolution and diversity order of four. However, the code rate achieved with the Tarokh code is half of the one achieved with the Alamouti code. Furthermore, the unambiguous range is reduced since the bandwidth is divided to multiplex the different antenna signals. For its simplicity, good performance and reduced integration requirements, the method is promising for future ISAC systems.

Removing False Targets for Cyclic Prefixed OFDM Sensing with Extended Ranging.

Employing a cyclic prefixed OFDM (CP-OFDM) communication waveform for sensing has attracted extensive attention in vehicular integrated sensing and communications (ISAC). A unified sensing framework was developed recently, enabling CP-OFDM sensing to surpass the conventional limits imposed by underlying communications. However, a false target issue still remains unsolved. In this paper, we investigate and solve this issue. Specifically, we unveil that false targets are caused by periodic cyclic prefixes (CPs) in CP-OFDM waveforms. We also derive the relation between the locations of false and true targets, and other features, e.g., strength, of false targets. Moreover, we develop an effective solution to remove false targets. Simulations are provided to confirm the validity of our analysis and the effectiveness of the proposed solution. In particular, our design can reduce the false alarm rate caused by false targets by over 50% compared with the prior art.

A Retrospective Clinical Audit of the ImmunoCAP ISAC 112 for Multiplex Allergen Testing.

INTRODUCTION: Complex cases of multiple allergies can be particularly difficult to diagnose using standard methods such as skin prick tests and assessment of a patient's allergic history. Multiplex allergy testing may improve outcomes for allergy patients by avoiding misdiagnosis and providing reassurance. The ImmunoCAP Immuno Solid-Phase Allergen Chip (ISAC) 112 is a CE-marked, molecular, multiplex, allergy test that can test for IgE antibodies to 112 components from 51 allergen sources. However, its clinical utility is unknown and is difficult to estimate due to the complexity of the diagnostic pathway in which it is used. OBJECTIVE: To assess how the ImmunoCAP ISAC 112 is currently being used in UK practice. The patient populations in which it may have the most benefit were examined, and the sequence of other tests implemented alongside ISAC was determined. METHODS: A retrospective audit of 100 patient cases from 2 UK tertiary allergy clinics was performed. Fifty paediatric and fifty adult cases were selected for audit. The indications for ordering an ISAC test, the other tests used alongside ISAC, and changes in management actioned by the ISAC test were investigated. RESULTS: 73.6% of paediatric and 78% of adult patients referred for an ISAC test were suspected to have multiple sensitizations. The sequence of testing varied greatly between cases, but 70% of adult and 98% of paediatric patients had at least one other investigation prior to an ISAC test. In most cases, ISAC testing confirmed clinical suspicion. CONCLUSIONS: A prospective research study is necessary to further investigate the clinical utility and cost-effectiveness of the ISAC. A UK national registry would be of great benefit but will require a large resource base.

Peanut components measured by ISAC: comparison with ImmunoCap and clinical relevance in peanut allergic children.

BACKGROUND: Specific IgE (sIgE) against the peanut component Arachis hypogaea (Ara h) 2 has been shown to be the most important allergen to discriminate between peanut allergy and peanut tolerance. Several studies determined sIgE cut off values for Ara h 2, determined by singleplex measurements. However, cut off values for Ara h 2 from multiplex arrays are less well defined. The aim of this study was to evaluate the correlation between Ara h 2 sIgE determined by singleplex versus multiplex measurements and to assess the diagnostic value of the different peanut components included in Immuno Solid-phase Allergen Chip (ISAC) multiplex analysis in children with a suspected peanut allergy. METHODS: In this retrospective study we analyzed Ara h 2 sIgE values with singleplex Fluorescence Enzyme Immunoassay (FEIA, ImmunoCap) and multiplex microarray (ISAC) measurements in 117 children with a suspected peanut allergy. Also, other peanut components measured by ISAC were analyzed. Double blinded placebo controlled oral food challenges were used as golden standard. RESULTS: Among all studied peanut components FEIA Ara h 2 sIgE showed the highest area under the curve (AUC, 0.922), followed by ISAC Ara h 6 and Ara h 2 sIgE with AUCs of respectively 0.906 and 0.902. Best cut off values to diagnose peanut allergy were 4.40 kU/l for FEIA Ara h 2 sIgE and, 7.43 ISU and 8.13 ISU for respectively Ara h 2 and Ara h 6 sIgE in ISAC microarray. Ara h 2 sIgE determined in FEIA and ISAC showed a good correlation (r = 0.88; p < 0.01). CONCLUSION: Ara h 6 and Ara h 2 sIgE in multiplex ISAC are both good predictors of clinical peanut allergy in Dutch children, and their performance is comparable to the use of Ara h 2 in singleplex FEIA. The simultaneous measurement of different peanut components using ISAC is an advantage and clinically useful to detect peanut allergic children that are Ara h 2 negative but sensitized to other peanut proteins such as Ara h 6.

Procedures for Flow Cytometry-Based Sorting of Unfixed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infected Cells and Other Infectious Agents.

In response to the recent COVID-19 pandemic, many laboratories are involved in research supporting SARS-CoV-2 vaccine development and clinical trials. Flow cytometry laboratories will be responsible for a large part of this effort by sorting unfixed antigen-specific lymphocytes. Therefore, it is critical and timely that we have an understanding of risk assessment and established procedures of infectious cell sorting. Here we present procedures covering the biosafety aspects of sorting unfixed SARS-CoV-2-infected cells and other infectious agents of similar risk level. These procedures follow the ISAC Biosafety Committee guidelines and were recently approved by the National Institutes of Health Institutional Biosafety Committee for sorting SARS-CoV-2-infected cells. © 2020 International Society for Advancement of Cytometry.