Gábor Transform-Based Antibody Quantitation in Serum: An Interlaboratory Liquid Chromatography/High-Resolution Mass Spectrometry Investigation.

Therapeutic monoclonal antibodies (t-mAbs) are crucial for treating various conditions, including cancers and autoimmune disorders. Accurate quantitation and pharmacokinetic monitoring of t-mAbs in serum are essential, but current methods like ligand binding assays (LBAs) and bottom-up peptide liquid chromatography-tandem mass spectrometry (LC-MS/MS) can lack the sensitivity and specificity needed to meet clinical demands. Emerging techniques using high-resolution mass spectrometry (HRMS) in top-down and middle-up approaches offer improved ability to accurately quantify mAb proteoforms apart from degradation products by keeping the sample proteins intact or minimizing digestion.

Experimental Insights into the Formation, Reactivity, and Crosstalk of Thionitrite (SNO) and Perthionitrite (SSNO).

Hydrogen sulfide (HS) and nitric oxide (NO) are important gaseous biological signaling molecules that are involved in complex cellular pathways. A number of physiological processes require both HS and NO, which has led to the proposal that different HS/NO⋅ crosstalk species, including thionitrite (SNO) and perthionitrite (SSNO), are responsible for this observed codependence. Despite the importance of these S/N hybrid species, the reported properties and characterization, as well as the fundamental pathways of formation and subsequent reactivity, remain poorly understood.

Strategies to mitigate electrostatic charging during coffee grinding.

Coffee grinding generates electrostatically charged particles, causing clumping, spark discharge, and beyond. When brewing, the particle aggregates affect liquid-solid surface accessibility, leading to variable extraction quality. Here, we study four charge mitigation strategies.

Bulking up: the impact of polymer sterics on emulsion stability.

Encapsulation of hydrophobic active ingredients is critical for targeted drug delivery as water-insoluble drugs dominate the pharmaceutical marketplace. We previously demonstrated hexadecane-in-water emulsions stabilized with a pH-tunable polymer, poly(acrylic acid) (PAA), a steric layer preventing particle aggregation. Using vibrational sum frequency scattering spectroscopy (VSFSS), here we probe the influence of steric hindrance on emulsion colloidal stability by tailoring the molecular weight of PAA and by adding an additional methyl group to the polymer backbone poly(methacrylic acid) (PMAA) at pH 2, 4, and 6.

Streamlining LC-MS Characterization of Pharmaceutical Polymers by Fourier-Transform-Based Deconvolution and Macromolecular Mass Defect Analysis.

Polymer conjugation has risen in importance over the past three decades as a means of increasing the half-life of biotherapeutics, with benefits including better stability, greater drug efficacy, and lower toxicity. However, the intrinsic variability of polymer synthesis results in products with broad distributions in chain length and branching structure, complicating quality control for successful functionalization and downstream conjugation. Frequently, a combination of several analytical techniques is required for comprehensive characterization.

Modeling collisional kinetic energy damping, heating, and cooling of ions in mass spectrometers: a tutorial perspective.

Many powerful methods in mass spectrometry rely on activation of ions by high-energy collisions with gas particles. For example, multiple Collision Induced Dissociation (CID) has been used for many years to determine structural information for ions ranging from small organics to large, native-like protein complexes. More recently, Collision Induced Unfolding (CIU) has proved to be a very powerful method for understanding high-order protein structure and detecting differences between similar proteins.

The complete assembly of human LAT1-4F2hc complex provides insights into its regulation, function and localisation.

The LAT1-4F2hc complex (SLC7A5-SLC3A2) facilitates uptake of essential amino acids, hormones and drugs. Its dysfunction is associated with many cancers and immune/neurological disorders. Here, we apply native mass spectrometry (MS)-based approaches to provide evidence of super-dimer formation (LAT1-4F2hc).

Reversible Hydrosulfide (HS) Binding Using Exclusively C-H Hydrogen-Bonding Interactions in Imidazolium Hosts.

HS is a physiologically important signaling molecule with complex roles in biology and exists primarily as HS at physiological pH. Despite this anionic character, few investigations have focused on the molecular recognition and reversible binding of this important biological anion. Using a series of imidazole and imidazolium host molecules, we investigate the role of preorganization and charge on HS binding.

Modulation of Carbon Nanotube Electronic Structure by Grain Boundary Defects in RbI on Au(111).

The electronic properties of single-walled carbon nanotubes (SWCNTs) are known to be highly sensitive to environmental effects. Here, we use scanning tunneling microscopy and spectroscopy to investigate the electronic properties of SWCNTs deposited on RbI monolayer films grown on Au(111). We find that grain boundary defects in RbI monolayers cause the appearance of spatially confined localized states in the SWCNTs.

Spatially modulated interface states in a two-dimensional potential: Single-layer RbI on Ag(111).

Alkali halides are known to exhibit interface electronic states (IES) when deposited on metal surfaces with ultra-thin coverage. Here, we examine the IES formed by sub-monolayer RbI growth on Ag(111), which exhibits spatial variations in electronic structure in surprising contrast to the results previously obtained for other alkali halides. We find that this spatially dependent behavior can be qualitatively modeled by using a two-dimensional cosine potential commensurate with the moiré superstructure, where the IES is constructed from the well-known analytical solutions to the Mathieu equation.

Eye lens β-crystallins are predicted by native ion mobility-mass spectrometry and computations to form compact higher-ordered heterooligomers.

Eye lens α- and β-/γ-crystallin proteins are not replaced after fiber cell denucleation and maintain lens transparency and refractive properties. The exceptionally high (∼400-500 mg/mL) concentration of crystallins in mature lens tissue and multiple other factors impede precise characterization of β-crystallin interactions, oligomer composition, size, and topology. Native ion mobility-mass spectrometry is used here to probe β-crystallin association and provide insight into homo- and heterooligomerization kinetics for these proteins.

Moderate-Affinity Affibodies Modulate the Delivery and Bioactivity of Bone Morphogenetic Protein-2.

Uncontrolled bone morphogenetic protein-2 (BMP-2) release can lead to off-target bone growth and other adverse events. To tackle this challenge, yeast surface display is used to identify unique BMP-2-specific protein binders known as affibodies that bind to BMP-2 with different affinities. Biolayer interferometry reveals an equilibrium dissociation constant of 10.

Structural Bistability in RbI Monolayers on Ag(111).

Alkali halides are well-known for their tendency to form rock-salt-like crystal structures. Here we present a scanning tunneling microscopy study of a previously unreported alternative structure of one such alkali halide, RbI. When deposited on Ag(111) at a low submonolayer surface coverage, RbI forms islands with hexagonally coordinated atomic structures, in contrast to the expected rock-salt structures typically observed for such alkali halide films on metal surfaces.

Investigation of Charge-State-Dependent Compaction of Protein Ions with Native Ion Mobility-Mass Spectrometry and Theory.

The precise relationship between native gas-phase protein ion structure, charge, desolvation, and activation remains elusive. Much evidence supports the Charge Residue Model for native protein ions formed by electrospray ionization, but scaling laws derived from it relate only to overall ion size. Closer examination of drift tube CCSs across individual native protein ion charge state distributions (CSDs) reveals deviations from global trends.

Lipid Head Group Adduction to Soluble Proteins Follows Gas-Phase Basicity Predictions: Dissociation Barriers and Charge Abstraction.

Native mass spectrometry analysis of membrane proteins has yielded many useful insights in recent years with respect to membrane protein-lipid interactions, including identifying specific interactions and even measuring binding affinities based on observed abundances of lipid-bound ions after collision-induced dissociation (CID). However, the behavior of non-covalent complexes subjected to extensive CID can in principle be affected by numerous factors related to gas-phase chemistry, including gas-phase basicity (GB) and acidity, shared-proton bonds, and other factors. A recent report from our group showed that common lipids span a wide range of GB values.

Solvent Cage Effects: A Comparison of Geminate and Nongeminate Radical Cage Pair Combination Efficiencies.

This study measured and compared the combination efficiencies () of geminate radical cage pairs to nongeminate (collisional) radical cage pairs ('). For the [Cp'(CO)Mo·, ·Mo(CO)Cp'] radical cage pair, ' was found to be smaller than in solutions having the same viscosity. It is proposed that the difference in and ' arises because the radicals in the collisional cage pair are less likely to have the correct orbital orientation for radical-radical combination to occur, whereas photochemically generated geminate cage pairs are more likely to have the correct orbital orientation.

Increasing Collisional Activation of Protein Complexes Using Smaller Aperture Source Sampling Cones on a Synapt Q-IM-TOF Instrument with a Stepwave Source.

Quadrupole ion mobility time-of-flight (Q-IM-TOF) mass spectrometers have revolutionized investigation of native biomolecular complexes. High pressures in the sources of these instruments aid transmission of protein complexes through damping of kinetic energy by collisional cooling. As adducts are removed through collisional heating (declustering), excessive collisional cooling can prevent removal of nonspecific adducts from protein ions, leading to inaccurate mass measurements, broad mass spectral peaks, and obfuscation of ligand binding.

Distinct classes of multi-subunit heterogeneity: analysis using Fourier Transform methods and native mass spectrometry.

Native electrospray mass spectrometry is a powerful method for determining the native stoichiometry of many polydisperse multi-subunit biological complexes, including multi-subunit protein complexes and lipid-bound transmembrane proteins. However, when polydispersity results from incorporation of multiple copies of two or more different subunits, it can be difficult to analyze subunit stoichiometry using conventional mass spectrometry analysis methods, especially when m/z distributions for different charge states overlap in the mass spectrum. It was recently demonstrated by Marty and co-workers (K.

Synthesis of Metastable Inorganic Solids with Extended Structures.

The number of known inorganic compounds is dramatically less than predicted due to synthetic challenges, which often constrains products to only the thermodynamically most stable compounds. Consequently, a mechanism-based approach to inorganic solids with designed structures is the holy grail of solid state synthesis. This article discusses a number of synthetic approaches using the concept of an energy landscape, which describes the complex relationship between the energy of different atomic configurations as a function of a variety of parameters such as initial structure, temperature, pressure, and composition.

Computational Insights into Compaction of Gas-Phase Protein and Protein Complex Ions in Native Ion Mobility-Mass Spectrometry.

Native ion mobility-mass spectrometry (IM-MS) is a rapidly growing field for studying the composition and structure of biomolecules and biomolecular complexes using gas-phase methods. Typically, ions are formed in native IM-MS using gentle nanoelectrospray ionization conditions, which in many cases can preserve condensed-phase stoichiometry. Although much evidence shows that large-scale condensed-phase structure, such as quaternary structure and topology, can also be preserved, it is less clear to what extent smaller-scale structure is preserved in native IM-MS.

Donor-/Acceptor-Substituted Tetrakis(arylethynyl)benzenes: The Influence of Donor Group on Optoelectronic Properties.

We have prepared nine structural isomers of a tetrakis(arylethynyl)benzene chromophore functionalized with 4-butoxyphenyl and pyridyl units as the respective donor and acceptor units and examined their steady-state spectroscopic parameters to study how small structural variations effect the electronic absorption and emission spectra. Unlike their 4-dibutylaminophenyl congeners that exhibited dynamic hypsochromic or bathochromic shifts in response to Lewis and Brønsted acids, the current class of compounds simply showed quenched fluorescence upon protonation; only AlCl elicited a red-shifted fluorescence response. Computational studies of each system were also performed to provide additional insight into the energy levels and electronic transitions present.

Serendipitous Rediscovery of the Facile Cyclization of Z,Z-3,5-Octadiene-1,7-diyne Derivatives to Afford Stable, Substituted Naphthocyclobutadienes.

The serendipitous isolation of very small amounts of two naphthocyclobutadiene (NCB) derivatives has led to the computational re-examination of the electrocyclization of Z,Z-3,5-octadiene-1,7-diyne as well as the experimental and computational study of diethynylindeno[2,1-a]fluorene derivatives that contain the 3,5-octadiene-1,7-diyne motif as part of a larger π-framework. In both cases the calculated potential energy surface strongly implicates two successive electrocyclic reactions to afford the antiaromatic products. With the octadienediyne fragment locked in the reactive conformation, the postulated diethynylindeno[2,1-a]fluorene intermediates afford the NCBs in modest to good yields.

On the Rise: Experimental and Computational Vibrational Sum Frequency Spectroscopy Studies of Pyruvic Acid and Its Surface-Active Oligomer Species at the Air-Water Interface.

It is well known that atmospheric aerosol play important roles in the environment. However, there is still much to learn about the processes that form aerosols, particularly aqueous secondary organic aerosols. While pyruvic acid (PA) is often better known for its biological significance, it is also an abundant atmospheric secondary organic ketoacid.

Takes Two to Tango: Choreography of the Coadsorption of CTAB and Hexanol at the Oil-Water Interface.

Mixed surfactant systems at the oil-water interface play a vital role in applications ranging widely from drug delivery to oil-spill remediation. Synergistic mixtures are superior emulsifiers and more effective at modifying surface tension than either component alone. Mixtures of surfactants with dissimilar polar head groups are of particular interest because of the additional degree of control they offer.