Search

A charge detector is a vital component in neutrino and dark matter detection. The integration of a charge collector in the form of flat pads and readout modules has been proposed as an optimization method as it can reduce noise and installation complexity. As a substrate, fused silica glass has attracted considerable attention due to its low radioactive background properties. In this research, based on the application requirements of a high charge collection rate and low noise, the structure of the charge detector was designed using calculation and simulation methods. The entire manufacturing process is described. In addition, a novel through glass via (TGV) structure composed of a conformal metal layer and a photosensitive material that is easy to fabricate and has high morphological compatibility with via filling is proposed. The curing property of the new material was characterized. A fully integrated solid-state charge detector with 32 groups of TGVs was realized. Additionally, the electrical properties of key structures were tested and analyzed.

Experiments on a modified Orbitrap Exploris instrument have shown that even in a compact instrument it is possible to achieve a resolving power in excess of 2,000,000 when appropriate tolerance and tuning requirements are met. Such levels are achieved for a 4-s detection time. A new Orbitrap design with improved pumping allowed, for the first time, isotopic resolution of intact monoclonal antibodies even when using standard nitrogen supply in the Ion Routing Multipole. Longer transients further improve signal-to-noise ratios, approaching levels needed for single-charge resolution in charge detection mass spectrometry. Further steps for improvement of instrumentation are discussed.

Charge detection mass spectrometry (CDMS) is a single-particle technique where the masses of individual ions are determined from simultaneous measurement of their mass-to-charge ratio (m/z) and charge. Masses are determined for thousands of individual ions, and then the results are binned to give a mass spectrum. Using this approach, accurate mass distributions can be measured for heterogeneous and high-molecular-weight samples that are usually not amenable to analysis by conventional mass spectrometry. Recent applications include heavily glycosylated proteins, protein complexes, protein aggregates such as amyloid fibers, infectious viruses, gene therapies, vaccines, and vesicles such as exosomes.

A new Orbitrap-based ion analysis procedure is shown to be possible by determining the direct charge for numerous individual protein ions to generate true mass spectra. The deployment of an Orbitrap system for charge detection enables the characterization of highly complicated mixtures of proteoforms and their complexes in both denatured and native modes of operation, revealing information not obtainable by traditional measurement of an ensemble of ions.

The masses of particles in a bovine milk extracellular vesicle (EV) preparation enriched for exosomes were directly determined for the first time by charge detection mass spectrometry (CDMS). In CDMS, both the mass-to-charge ratio (m/z) and z are determined simultaneously for individual particles, enabling mass determinations for particles that are far beyond the mass limit (∼1.0 MDa) of conventional mass spectrometry (MS). Particle masses and charges span a wide range from m ∼ 2 to ∼90 MDa and z ∼ 50 to ∼1300 e (elementary charges) and are highly dependent upon the conditions used to extract and isolate the EVs. EV particles span a continuum of masses, reflecting the highly heterogeneous nature of these samples. However, evidence for unique populations of particles is obtained from correlation of the charges and masses. An analysis that uses a two-dimensional Gaussian model, provides evidence for six families of particles, four of which having masses in the range expected for exosomes. Complementary proteomics measurements and electron microscopy (EM) imaging are used to further characterize the EVs and confirm that these samples have been enriched in exosomes. The ability to characterize such extremely heterogeneous mixtures of large particles with rapid, sensitive, and high-resolution MS techniques is critical to ongoing analytical efforts to separate and purify exosomes and exosome subpopulations. Direct measurement of each particle's mass and charge is a new means of characterizing the physical and chemical properties of exosomes and other EVs.

Virus-like particle (VLP) conjugates are being developed for biomedical applications; however, there is a lack of quantitative analytical methods to measure the extent of conjugation and modification of VLP based therapeutics. Charge detection mass spectrometry (CDMS) can measure mass distributions for large and heterogeneous complexes and is emerging as a valuable tool in the analysis of biologics. In this study, CDMS is used to characterize the stoichiometry and population distribution of antibodies covalently conjugated to the surface of a bacteriophage MS2 VLP. Initial CDMS analysis of the unconjugated MS2 particles suggested that they had packaged a broad distribution of exogenous genomic material. We developed procedures to remove the undesired genomic material from the VLP preparation and observed that, for the samples where the genomic fragments were removed, the antibody coupling reaction efficiency increased by almost a factor of 2. This meant there were (1) fewer VLPs with no antibodies bound, which is an important consideration for the efficacy of a targeted therapeutic and (2) fewer antibodies were wasted during the coupling reaction. CDMS could be employed in a similar manner as a tool to characterize coupling reaction product distributions and precursors and help inform the development of the next generation of conjugate-based therapies.

Adeno-associated virus (AAV) vectors are clinically proven gene delivery vehicles that are attracting an increasing amount of attention. Non-genome-containing empty AAV capsids are by-products during AAV production that have been reported to potentially impact AAV product safety and efficacy. Therefore, the presence and amount of empty AAV capsids need to be characterized during process development. Multiple methods have been reported to characterize empty AAV capsid levels, including transmission electron microscopy (TEM), analytical ultracentrifugation (AUC), charge detection mass spectrometry (CDMS), UV spectrophotometry, and measuring capsid and genome copies by ELISA and qPCR. However, these methods may lack adequate accuracy and precision or be challenging to transfer to a quality control (QC) lab due to the difficulty of implementation. In this study, we used AAV serotype 6.2 (AAV6.2) as an example to show the development of a QC-friendly anion exchange chromatography (AEX) assay for the determination of empty and full capsid percentages. The reported assay requires several microliters of material with a minimum titer of 5 × 1011 vg/mL, and it can detect the presence of as low as 2.9% empty capsids in AAV6.2 samples. Additionally, the method is easy to deploy, can be automated, and has been successfully implemented to support testing of various in-process and release samples. Keywords: AAV, AAV6.2, Chromatography, Anion exchange chromatography (AEC), Empty capsids, AUC, High-throughput

Native mass spectrometry (MS) has become a versatile tool for characterizing high-mass complexes and measuring biomolecular interactions. Native MS usually requires resolution of different charge states produced by electrospray ionization to measure the mass, which is difficult for highly heterogeneous samples that have overlapping and unresolvable charge states. Charge detection-mass spectrometry (CD-MS) seeks to address this challenge by simultaneously measuring the charge and m/z for isolated ions. However, CD-MS often shows uncertainty in the charge measurement that limits the resolution. To overcome this charge state uncertainty, we developed UniDecCD (UCD) software for computational de-convolution of CD-MS data, which significantly improves the resolution of CD-MS data. Here, we describe the UCD algorithm and demonstrate its ability to improve CD-MS resolution of proteins, megadalton viral capsids, and heterogeneous nanodiscs made from natural lipid extracts. UCD provides a user-friendly interface that will increase the accessibility of CD-MS technology and provide a valuable new computational tool for CD-MS data analysis.

Adeno-associated viruses (AAV) represent important gene therapy vectors with several approved clinical applications and numerous more in clinical trials. Genome packaging is an essential step in the bioprocessing of AAVs and needs to be tightly monitored to ensure the proper delivery of transgenes and the production of effective drugs. Current methods to monitor genome packaging have limited sensitivity, a high demand on labour, and struggle to distinguish between packaging of the intended genome or unwanted side-products. Here we show that Orbitrap based charge detection mass spectrometry allows the ultra-sensitive quantification of all these different AAV bioprocessing products. A protocol is presented that allows the quantification of genome packed AAV preparations in under half an hour, requiring only micro-liter quantities of typical AAV preparations with ~1013 viral genome copies per millilitre. The method quickly assesses the integrity and amount of genome packed AAV particles to support AAV bioprocessing and characterization of this rapidly emerging class of advanced drug therapies.

We report on charge detection in electrostatically-defined quantum dot devices in bilayer graphene using an integrated charge detector. The device is fabricated without any etching and features a graphite back gate, leading to high quality quantum dots. The charge detector is based on a second quantum dot separated from the first dot by depletion underneath a 150 nm wide gate. We show that Coulomb resonances in the sensing dot are sensitive to individual charging events on the nearby quantum dot. The potential change due to single electron charging causes a step-like change (up to 77 %) in the current through the charge detector. Furthermore, the charging states of a quantum dot with tunable tunneling barriers and of coupled quantum dots can be detected.

Applications of charge detection mass spectrometry (CDMS) for measuring the masses of large molecules, macromolecular complexes, and synthetic polymers that are too large or heterogeneous for conventional mass spectrometry measurements are made possible by weighing individual ions in order to avoid interferences between ions. Here, a new multiplexing method that makes it possible to measure the masses of many ions simultaneously in CDMS is demonstrated. Ions with a broad range of kinetic energies are trapped. The energy of each ion is obtained from the ratio of the intensity of the fundamental to the second harmonic frequencies of the periodic trapping motion making it possible to measure both the m/ z and charge of each ion. Because ions with the exact same m/ z but with different energies appear at different frequencies, the probability of ion-ion interference is significantly reduced. We show that the measured mass of a protein complex consisting of 16 protomers, RuBisCO (517 kDa), is not affected by the number of trapped ions with up to 21 ions trapped simultaneously in these experiments. Ion-ion interactions do not affect the ion trapping lifetime up to 1 s, and there is no influence of the number of ions on the measured charge-state distribution of bovine serum albumin (66.5 kDa), indicating that ion-ion interactions do not adversely affect any of these measurements. Over an order of magnitude gain in measurement speed over single ion analysis is demonstrated, and significant additional gains are expected with this multi-ion measurement method.

Reading out Majorana bound states (MBSs) is essential both to verify their non-Abelian property and to realize topological quantum computation. Here, we construct a protocol to measure the parity of two MBSs in a Majorana island coupled to double quantum dot (DQD). The parity information is mapped to the charge state of the DQD through Landau-Zener transition. The operation needed is sweeping the bias of the DQD, which is followed by charge sensing. In the case without fine-tuning, a single run of sweep-and-detection implement a weak measurement of the parity. We find that in general a sequence of about ten runs would completely project a superposition state to either parity, and the charge detection in each run records how the state of MBSs collapses step by step. Remarkably, this readout protocol is of non-demolition and robust to low frequency charge fluctuation., 7 pages, 5 figures

Adeno-associated viruses (AAVs) represent important gene therapy vectors with several approved clinical applications and numerous more in clinical trials. Genome packaging is an essential step in the bioprocessing of AAVs and needs to be tightly monitored to ensure the proper delivery of transgenes and the production of effective drugs. Current methods to monitor genome packaging have limited sensitivity, a high demand on labor, and struggle to distinguish between packaging of the intended genome or unwanted side-products. Here we show that Orbitrap-based charge-detection mass spectrometry allows the very sensitive quantification of all these different AAV bioprocessing products. A protocol is presented that allows the quantification of genome-packed AAV preparations in under half an hour, requiring only micro-liter quantities of typical AAV preparations with ∼1013 viral capsids per milliliter. The method quickly assesses the integrity and amount of genome packed AAV particles to support AAV bioprocessing and characterization of this rapidly emerging class of advanced drug therapies., Graphical abstract, Orbitrap-based charge-detection mass spectrometry can efficiently assess the integrity and amount of genome-packed AAV particles, supporting the bioprocessing and characterization of this emerging class of advanced drug products.

The ability of field effect transistors (FETs) to detect charge variations on the gate may be exploited for realizing chemo- and bio-sensors. In this paper, we focus our attention on a particular kind of field effect device, named organic charge modulated FETs, whose features can be optimized for charge detection in liquid solutions. The results of the measurement of different bio-related effects are shown. In particular, DNA hybridization and pH detection in liquid media are proposed. Finally, preliminary considerations about the applicability of these devices to the detection of the electrical activity of cells are also provided. The device has considerable potential for being employed as a reliable, high sensitivity, low cost technology for sensing signals derived from living systems. This journal is © 2013 The Royal Society of Chemistry.

During the development of new battery technologies, high production tolerances are likely to occur due to the number of manual manufacturing steps. When putting these prototypes into operation, one of the most critical parameters is the reliable state of charge detection. This can be challenging when parameters like the capacity or the end of charge voltage are not precisely known due to the above-mentioned tolerances. In the majority of cases overcharging should be avoided as it harms the battery. This paper proposes a new criterion for detecting the end of the charging process that is based on the rate of change of electrochemical impedance spectra of the examined batteries. Device parameter fluctuations influence every measurement. Therefore, using the rate of change offers the advantage of using relative values instead of absolute values.

Lipoproteins are micelle-like assemblies that are key players in the pathogenesis of atherosclerosis. High-density lipoprotein (HDL), low-density lipoproteins (LDL), and very low density lipoprotein (VLDL) are the three major classes present in fasting plasma. Within each class, there is a broad size distribution with wide variations in protein and lipid content. The development of better metrics for cardiovascular risk is thought to depend on better characterization of lipoprotein subclasses. Using charge detection mass spectrometry (CDMS), the mass distributions of HDL, LDL, and VLDL have been directly measured for the first time. In the case of HDL, seven distinct subpopulations were resolved using a two-dimensional correlation of charge and mass. The resolved components are assigned to HDL particles containing different numbers of the key structural proteins apolipoprotein A-I and apolipoprotein A-II.

International audience; Heterogeneity and polymorphism are generic features of amyloid fibers with some important effects on the related disease development. We report here the characterization, by charge detection mass spectrometry, of amyloid fibers made of three polypeptides involved in neurodegenerative diseases: Aβ1–42 peptide, tau and α-synuclein. Beside the mass of individual fibers, this technique enables to characterize the heterogeneity and the polymorphism of the population. In the case of Aβ1–42 peptide and tau protein, several coexisting species could be distinguished and characterized. In the case of α-synuclein, we show how the polymorphism affects the mass and charge distributions.

Hepatitis B virus (HBV) core protein is a model system for studying assembly and disassembly of icosahedral structures. Controlling disassembly will allow re-engineering the 120 subunit HBV capsid, making it a molecular breadboard. We examined removal of subunits from partially crosslinked capsids to form stable incomplete particles. To characterize incomplete capsids, we used two single molecule techniques, resistive-pulse sensing and charge detection mass spectrometry. We expected to find a binomial distribution of capsid fragments. Instead, we found a preponderance of 3 MDa complexes (90 subunits) and no fragments smaller than 3 MDa. We also found 90-mers in the disassembly of uncrosslinked HBV capsids. 90-mers seem to be a common pause point in disassembly reactions. Partly explaining this result, graph theory simulations have showed a threshold for capsid stability between 80 and 90 subunits. To test a molecular breadboard concept, we showed that missing subunits could be refilled resulting in chimeric, 120 subunit particles. This result may be a means of assembling unique capsids with functional decorations. This article is protected by copyright. All rights reserved.

Spontaneous mass and charge losses from individual multi-megadalton ions have been observed with charge detection mass spectrometry (CDMS) by trapping single hepatitis B virus (HBV) capsids for 3 s. Gradual increases in the oscillation frequency of single ions in the ion trap are attributed mainly to mass loss (probably solvent, water, and/or salt). The total mass lost during the 3 s trapping period peaks at around 20 kDa for 4 MDa HBV T = 4 capsids. Discrete frequency drops punctuate the gradual increases in the oscillation frequencies. The drops are attributed to a sudden loss of charge. In most cases a single positive charge is lost along with some mass (on average around 1000 Da). Charge loss occurs for over 40% of the trapped ions. It usually occurs near the beginning of the trapping event, and it occurs preferentially in regions of the trap with strong electric fields, indicating that external electric fields promote charge loss. This process may contribute to the decrease in m/z resolution that often occurs with megadalton ions. Graphical Abstract ᅟ.

G-protein Coupled Receptors (GPCRs) form the largest membrane protein superfamily in vertebrates. Advances in crystallization techniques so far resulted in the resolution of 44 unique receptors available for the GPCRs researcher’s community, 37 of which belong to rhodopsin-like GPCRs class. We performed here the first systematic analysis of GPCRs binding cavities based on the available pool of rhodopsin-like solved structures. We pinpointed ten positions shared between all the solved receptors, namely 3.32, 3.33, 3,36, 6.48, 6.51, 6.52, 6.55, 7.35, 7.39 and 7.43, as interacting with ligands. We analyzed the conservation of amino acids present in these positions and clustered GPCRs accordingly to the physicochemical properties of binding cavities’ residues. Clustering supplied new interesting insights into the common binding mode of these receptors. In particular, the 3.32 position turned out to have an important role in ligand charge detection. Finally, we demonstrated that residues in these ten positions have co-evolved together, sharing a common evolutionary history.

Charge detection mass spectrometry (CDMS) is a single molecule method where the mass of each ion is directly determined from individual measurements of its mass-to-charge ratio and charge. CDMS is particularly valuable for the analysis of high mass and heterogeneous analytes, where conventional MS methods are often confounded. In the last few years, CDMS has received a renaissance. Technical developments have improved the resolution and dramatically increased the breadth of problems that can be addressed. These improvements have moved CDMS more into the mainstream as interest in the application of mass spectrometry to high molecular weight species has grown. In the article, the three main variants of CDMS are described, along with an overview of recent applications.

We describe the design and principle of operation of a fast and sensitive electrometer operated at millikelvin temperatures, which aims at replacing conventional semiconducting charge amplifiers in experiments needing low back-action or high sensitivity. The electrometer consists of a Cooper Pair box (CPB) coupled to a microwave resonator, which converts charge variations to resonance frequency shifts. We analyze the dependence of the sensitivity on the various parameters of the device, and derive their optimization. By exploiting the nonlinearities of this electrometer, and using conventional nanofabrication and measurement techniques, a charge sensitivity of a few $10^{- 7} e / \sqrt{Hz}$ can be achieved which outperforms existing single charge electrometers., 13 pages, 7 figures

In this work, we report a new design of a charge detection quadrupole ion trap mass spectrometer (QIT-MS) for the analysis of micro-sized dry inorganic and bioparticles including red blood cells (RBCs) and different sizes of MCF-7 breast cancer cells. The developed method is one of the fastest methods to measure the mass of micro-sized particles. This system allows the online analysis of various micro-sized particles up to 1 x 10(17) Da. The calibration of the mass spectrometer has been done by using different sizes of polystyrene (PS) particles (2-15 mu m). The measured masses of RBCs were around 1.8 x 10(13) Da and MCF-7 cancer cells were between 1 x 10(14) and 4 x 10(14) Da. The calculated mass distribution profiles of the particles and cells were given as histogram profiles. The statistical data were summarized after Gaussian type fitting to the experimental histogram profiles. The new method gives very promising results for the analysis of particles and has very broad application.

Mass spectrometry has traditionally been the technology of choice for small molecule analysis, making significant inroads into metabolism, clinical diagnostics, and pharmacodynamics since the 1960s. In the mid-1980s, with the discovery of electrospray ionization (ESI) for biomolecule analysis, a new door opened for applications beyond small molecules. Initially, proteins were widely examined, followed by oligonucleotides and other nonvolatile molecules. Then in 1991, three intriguing studies reported using mass spectrometry to examine noncovalent protein complexes, results that have been expanded on for the last 25 years. Those experiments also raised the questions: How soft is ESI, and can it be used to examine even more complex interactions? Our lab addressed these questions with the analyses of viruses, which were initially tested for viability following electrospray ionization and their passage through a quadrupole mass analyzer by placing them on an active medium that would allow them to propagate. This observation has been replicated on multiple different systems, including experiments on an even bigger microbe, a spore. The question of analysis was also addressed in the early 2000s with charge detection mass spectrometry. This unique technology could simultaneously measure mass-to-charge and charge, allowing for the direct determination of the mass of a virus. More recent experiments on spores and enveloped viruses have given us insight into the range of mass spectrometry's capabilities (reaching 100 trillion Da), beginning to answer fundamental questions regarding the complexity of these organisms beyond proteins and genes, and how small molecules are integral to these supramolecular living structures. Graphical Abstract ᅟ.

The simulated noise used to benchmark wavelet edge detection in this work was described incorrectly. The correct description is given here, and new results based on noise that matches the original description are provided. The results support our original conclusion, which is that wavelet edge detection outperforms thresholding in the presence of white noise and 1/ f noise., (© 2021 IOP Publishing Ltd.)

Enabling fast charge of lithium-ion batteries is an essential step towards the wider adoption of electric vehicles. A key challenge is that high charging currents can promote several aging mechanisms, including lithium plating on the graphite anode. There is research suggesting that the deposition of metallic lithium favors further Li plating in a self-reinforcing loop, which can eventually lead to sudden cell failure.[1] Hence, detecting Li deposition during the early stages and developing electrochemical cycling protocols that avoid plating conditions are crucial to enabling fast-charge. In this talk we will discuss two approaches to investigate Li deposition. The first one is through detection. Our group has demonstrated that plated Li can intensify the Raman signal of its surrounding SEI, which then serves as an indirect marker for the presence of Li nuclei.[2] The sensitivity of this technique enables the study of where Li will tend to plate, and how this deposition evolves spatially over the course of many cycles. The second approach is through the use of a reference electrode, which can be used to estimate electrode potentials during fast charging.[3] This capability supports the development of fast-charging protocols that optimize time utilization while steering the cells away from Li plating conditions. Examples of such cycling protocols will be discussed during the presentation. References [1] X.-G. Yang, Y. Leng, G. Zhang, S. Ge, C.-Y. Wang, Journal of Power Sources, 360, 28-40 (2017) [2] M.-T. F. Rodrigues, V. A. Maroni, D. J. Gosztola, K. P. C. Yao, K. Kalaga, I. A. Shkrob, and D. P. Abraham, ACS Applied Energy Materials, 2 (1), 873-881 (2019) [3] M.-T. F. Rodrigues, K. Kalaga, S. E. Trask, D. W. Dees, I. A. Shkrob, and D. P. Abraham, Journal of The Electrochemical Society, 166 (6), A996-A1003 (2019) Acknowledgements: We are grateful to the U.S. DOE Office of Vehicle Technology Extreme Fast Charge Cell Evaluation of Lithium-Ion Batteries (XCEL) Program managed by Samuel Gillard. This document has been created by UChicago Argonne, LLC, Operator of Argonne National Laboratory (“Argonne”). Argonne, a U.S. Department of Energy Office of Science laboratory, is operated under Contract No. DE-AC02-06CH11357.

We demonstrate single-particle charge detection mass spectrometry on an Orbitrap for the analysis of megadalton biomolecular assemblies. We establish that the signal amplitudes of individual ions scale linearly with their charge, which can be used to resolve mixed ion populations, determine charge states and thus also determine the masses of individual ions. This enables the ultrasensitive analysis of heterogeneous protein assemblies including immunoglobulin oligomers, ribosomes, proteinaceous nanocontainers and genome-packed adeno-associated viruses.