Development of broad modulus profile upon polymer-polymer interface formation between immiscible glassy-rubbery domains.

Interfaces of glassy materials such as thin films, blends, and composites create strong unidirectional gradients to the local heterogeneous dynamics that can be used to elucidate the length scales and mechanisms associated with the dynamic heterogeneity of glasses. We focus on bilayer films of two different polymers with very different glass transition temperatures ([Formula: see text]) where previous work has demonstrated a long-range (∼200 nm) profile in local [Formula: see text] is established between immiscible glassy and rubbery polymer domains when the polymer-polymer interface is formed to equilibrium. Here, we demonstrate that an equally long-ranged gradient in local modulus [Formula: see text] is established when the polymer-polymer interface ([Formula: see text]5 nm) is formed between domains of glassy polystyrene (PS) and rubbery poly(butadiene) (PB), consistent with previous reports of a broad [Formula: see text] profile in this system. A continuum physics model for the shear wave propagation caused by a quartz crystal microbalance across a PB/PS bilayer film is used to measure the viscoelastic properties of the bilayer during the evolution of the PB/PS interface showing the development of a broad gradient in local modulus [Formula: see text] spanning [Formula: see text]180 nm between the glassy and rubbery domains of PS and PB. We suggest these broad profiles in [Formula: see text] and [Formula: see text] arise from a coupling of the spectrum of vibrational modes across the polymer-polymer interface as a result of acoustic impedance matching of sound waves with [Formula: see text] nm during interface broadening that can then trigger density fluctuations in the neighboring domain.

Quartz crystal microbalance-based aptasensor integrated with magnetic pre-concentration system for detection of Listeria monocytogenes in food samples.

A fast and accurate identification of Listeria monocytogenes. A new quartz crystal microbalance (QCM) aptasensor was designed for the specific and rapid detection of L. monocytogenes. Before detection of the target bacterium from samples in the QCM aptasensor, a magnetic pre-enrichment system was used to eliminate any contaminant in the samples. The prepared magnetic system was characterized using ATR-FTIR, SEM, VSM, BET, and analytical methods. The saturation magnetization values of the Fe3O4, Fe3O4@PDA, and Fe3O4@PDA@DAPEG particles were 57.2, 40.8, and 36.4 emu/g, respectively. The same aptamer was also immobilized on the QCM crystal integrated into QCM flow cell and utilized to quantitatively detect L. monocytogenes cells from the samples. It was found that a specific aptamer-magnetic pre-concentration system efficiently captured L. monocytogenes cells in a short time (approximately 10 min). The Fe3O4@PDA@DA-PEG-Apt particles provided selective isolation of L. monocytogenes from the bacteria-spiked media up to 91.8%. The immobilized aptamer content of the magnetic particles was 5834 µg/g using 500 ng Apt/mL. The QCM aptasensor showed a very high range of analytical performance to the target bacterium from 1.0 × 102 and 1.0 × 107 CFU/mL. The limit of detection (LOD) and limit of quantitation (LOQ) were 148 and 448 CFU/mL, respectively, from the feeding of the QCM aptasensor flow cell with the eluent of the magnetic pre-concentration system. The reproducibility of the aptasensor was more than 95%. The aptasensor was very specific to L. monocytogenes compared to the other Listeria species (i.e., L. ivanovii, L. innocua, and L. seeligeri) or other tested bacteria such as Staphylococcus aureus, Escherichia coli, and Bacillus subtilis. The QCM aptasensor was regenerated with NaOH solution, and the system was reused many times.

Piezoelectric biosensors: shedding light on principles and applications.

The three decades of experience with piezoelectric devices applied in the field of bioanalytical chemistry are shared. After introduction to principles and suitable measuring approaches, active and passive methods based on oscillators and impedance analysis, respectively, the focus is directed towards biosensing approaches. Immunosensing examples are provided, followed by other affinity sensing approaches based on hybridization of nucleic acids, aptamers, monitoring of enzyme activities, and detection of pathogenic microbes. The combination of piezosensors with cell lines and testing of drugs is highlighted, including mechanically active cells. The combination of piezosensors with other measuring techniques providing original hybrid devices is briefly discussed.

Influence of Front-End Electronics on Metrological Performance of QCM Systems.

Quartz Crystal Microbalances (QCMs) are versatile sensors employed in various fields, from environmental monitoring to biomedical applications, owing mainly to their very high sensitivity. However, the assessment of their metrological performance, including the impact of conditioning circuits, digital processing algorithms, and working conditions, is a complex and novel area of study. The purpose of this work is to investigate and understand the measurement errors associated with different QCM measurement techniques, specifically focusing on the influence of conditioning electronic circuits. Through a tailored and novel experimental setup, two measurement architectures-a Quartz Crystal Microbalance with dissipation monitoring (QCM-D) system and an oscillator-based QCM-R system-were compared under the same mechanical load conditions. Through rigorous experimentation and signal processing techniques, the study elucidated the complexities of accurately assessing QCM parameters, especially in liquid environments and under large mechanical loads. The comparison between the two different techniques allows for highlighting the critical aspects of the measurement techniques. The experimental results were discussed and interpreted based on models allowing for a deep understanding of the measurement problems encountered with QCM-based measurement systems. The performance of the different techniques was derived, showing that while the QCM-D technique exhibited higher accuracy, the QCM-R technique offered greater precision with a simpler design. This research advances our understanding of QCM-based measurements, providing insights for designing robust measurement systems adaptable to diverse conditions, thus enhancing their effectiveness in various applications.

Development of Nanopackaging for Storage and Transport of Loaded Lipid Nanoparticles.

Easily deploying new vaccines globally to combat disease outbreaks has been highlighted as a major necessity by the World Health Organization. RNA-based vaccines using lipid nanoparticles (LNPs) as a drug delivery system were employed to great effect during the recent COVID-19 pandemic. However, LNPs are still unstable at room temperature and agglomerate over time during storage, rendering them ineffective for intracellular delivery. We demonstrate the suitability of nanohole arrays (nanopackaging) as patterned surfaces to separate and store functionalized LNPs (fLNPs) in individual recesses, which can be expanded to other therapeutics. Encapsulating calcein as a model drug, we show through confocal microscopy the effective loading of fLNPs into our nanopackaging for both wet and dry systems. We prove quantifiably pH-mediated capture and subsequent unloading of over 30% of the fLNPs using QCM-D on alumina surfaces altering the pH from 5.5 to 7, displaying controllable storage at the nanoscale.

Cholesterol-Containing Liposomes Decorated With Au Nanoparticles as Minimal Tunable Fusion Machinery.

Membrane fusion is essential for the basal functionality of eukaryotic cells. In physiological conditions, fusion events are regulated by a wide range of specialized proteins, operating with finely tuned local lipid composition and ionic environment. Fusogenic proteins, assisted by membrane cholesterol and calcium ions, provide the mechanical energy necessary to achieve vesicle fusion in neuromediator release. Similar cooperative effects must be explored when considering synthetic approaches for controlled membrane fusion. We show that liposomes decorated with amphiphilic Au nanoparticles (AuLips) can act as minimal tunable fusion machinery. AuLips fusion is triggered by divalent ions, while the number of fusion events dramatically changes with, and can be finely tuned by, the liposome cholesterol content. We combine quartz-crystal-microbalance with dissipation monitoring (QCM-D), fluorescence assays, and small-angle X-ray scattering (SAXS) with molecular dynamics (MD) at coarse-grained (CG) resolution, revealing new mechanistic details on the fusogenic activity of amphiphilic Au nanoparticles (AuNPs) and demonstrating the ability of these synthetic nanomaterials to induce fusion regardless of the divalent ion used (Ca2+ or Mg2+ ). The results provide a novel contribution to developing new artificial fusogenic agents for next-generation biomedical applications that require tight control of the rate of fusion events (e.g., targeted drug delivery).

Selection of Highly Specific DNA Aptamer for the Development of QCM-Based Arsenic Sensor.

Heavy metal arsenic is a water pollutant that affects millions of lives worldwide. A novel aptamer candidate for specific and sensitive arsenic detection was identified using Graphene Oxide-SELEX (GO-SELEX). Eleven rounds of GO-SELEX were performed to screen As(III) specific sequences. The selected aptamer sequences were evaluated for their binding affinity. The dissociation constant of the best aptamer candidate, As-06 was estimated by fluorescence recovery upon target addition, and it was found to be 8.15 nM. A QCM-based biosensing platform was designed based on the target-triggered release of aptamer from the QCM electrode. An rGO-SWCNT nanocomposite was adsorbed on the gold surface, and the single-stranded probe was stacked on the rGO-CNT layer. Upon addition of the target to the solution, a concentration-dependent release of the ssDNA probe was observed and recorded as the change in the electrode frequency. The developed QCM sensor showed a dynamic linear range from 10 nM to 100 nM and a low detection limit of 8.6 nM. The sensor exhibited excellent selectivity when challenged with common interfering anions and cations.

A quartz crystal microbalance biosensor based on polyethylenimine-modified gold electrode to detect hepatitis B biomarker.

Biomarkers-based QCM-biosensors are suitable tools for the label-free detection of infectious diseases. In the current study, a QCM-biosensor was developed for the detection of HBsAg. Briefly, anti-HBsAg antibodies were covalently bound to the primary amines after PEI and thiolated-PEI surface modifications of gold-electrode. After RSM optimization, the statistical analysis revealed no significant difference between the immobilization yields of modified layers. Therefore, the PEI-modified QCM-biosensor was selected for further analysis. The PEI-surface was evaluated by FESEM, AFM, ATR-FTIR, and CA measurement. The surface hydrophilicity and its roughness were increased after PEI-coating. Also, FTIR confirmed the PEI-layering on the gold-surface. RSM optimization increased the antibody immobilization yield up to 80%. The QCM-biosensor showed noteworthy results with a wide dynamic range of 1-1 × 103 ng/mL, LOD of 3.14 ng/mL, LOQ of 9.52 ng/mL, and detection capability in human-sera, which were comparable with the ELISA. The mean accuracy of the QCM-biosensor was obtained at 91% when measured by the spike recovery test using human-sera. The biosensor was completely regenerated using 50 mM NaOH and 1% SDS. The benefits provided by the developed biosensor such as broad dynamic range, sensitivity, selectivity, stability, regenerate ability, and low cost suggest its potential application for the non-invasive and timely monitoring of HBV-biomarker.

Engineering and characterization of carbohydrate-binding modules for imaging cellulose fibrils biosynthesis in plant protoplasts.

Carbohydrate binding modules (CBMs) are noncatalytic domains that assist tethered catalytic domains in substrate targeting. CBMs have therefore been used to visualize distinct polysaccharides present in the cell wall of plant cells and tissues. However, most previous studies provide a qualitative analysis of CBM-polysaccharide interactions, with limited characterization of engineered tandem CBM designs for recognizing polysaccharides like cellulose and limited application of CBM-based probes to visualize cellulose fibrils synthesis in model plant protoplasts with regenerating cell walls. Here, we examine the dynamic interactions of engineered type-A CBMs from families 3a and 64 with crystalline cellulose-I and phosphoric acid swollen cellulose. We generated tandem CBM designs to determine various characteristic properties including binding reversibility toward cellulose-I using equilibrium binding assays. To compute the adsorption (nkon ) and desorption (koff ) rate constants of single versus tandem CBM designs toward nanocrystalline cellulose, we employed dynamic kinetic binding assays using quartz crystal microbalance with dissipation. Our results indicate that tandem CBM3a exhibited the highest adsorption rate to cellulose and displayed reversible binding to both crystalline/amorphous cellulose, unlike other CBM designs, making tandem CBM3a better suited for live plant cell wall biosynthesis imaging applications. We used several engineered CBMs to visualize Arabidopsis thaliana protoplasts with regenerated cell walls using confocal laser scanning microscopy and wide-field fluorescence microscopy. Lastly, we also demonstrated how CBMs as probe reagents can enable in situ visualization of cellulose fibrils during cell wall regeneration in Arabidopsis protoplasts.

Mycotoxins detection: view in the lens of molecularly imprinted polymer and nanoparticles.

Molecularly imprinted polymers (MIPs) are tailor-made functional composites which selectively recognize and bind the target molecule of interest. MIP composites are products of the massively cross-linked polymer matrices, generated via polymerization, with bio-inspired recognition cavities that are morphologically similar in size, shape and spatial patterns to the target conformation. These features have enabled researchers to expand the field of molecular recognition, more specifically for target with peculiar requirements. Nevertheless, MIPs alone are characterized with weak sensitivity. Besides, nanoparticles (NPs) are remarkably sensitive but also suffer from poor selectivity. Intriguingly, the combination of the two results in a highly sensitive and selective MIP composite. For instance, the conjugation of different functional NPs with MIPs can generate new flexible target capture tools, either a dynamic sensor or a novel drug delivery system. In this regard, although the technology is considered an established and feasible approach, it is still perceived as a burgeoning technology for various fields, which makes it unceasingly worthy reviewing. Therefore, in this review, we attempt to give an update on various custom-made biosensors based on MIPs in combination with various NPs for the detection of mycotoxins, the toxic secondary metabolites of fungi. We first summarize the classification, prevalence, and toxicological characteristics of common mycotoxins. Next, we provide an overview of MIP composites and their characterization, and then segment the role of NPs with respect to common types of MIP-based sensors. At last, conclusions and outlook are discussed.

Significance of Co-ion Partitioning in Salt Transport through Polyamide Reverse Osmosis Membranes.

Salt permeability of polyamide reverse osmosis (RO) membranes has been shown to increase with increasing feed salt concentration. The dependence of salt permeability on salt concentration has been attributed to the variation of salt partitioning with feed salt concentration. However, studies using various analytical techniques revealed that the salt (total ion) partitioning coefficient decreases with increasing salt concentration, in marked contrast to the observed increase in salt permeability. Herein, we thoroughly investigate the dependence of total ion and co-ion partitioning coefficients on salt concentration and solution pH. The salt partitioning is measured using a quartz crystal microbalance (QCM), while the co-ion partitioning is calculated from the measured salt partitioning using a modified Donnan theory. Our results demonstrate that the co-ion and total ion partitioning behave entirely differently with increasing salt concentrations. Specifically, the co-ion partitioning increased fourfold, while total ion partitioning decreased by 60% as the salt (NaCl) concentration increased from 100 to 800 mM. The increase in co-ion partitioning with increasing salt concentration is in accordance with the increasing trend of salt permeability in RO experiments. We further show that the dependence of salt and co-ion partitioning on salt concentration is much more pronounced at a higher solution pH. The good co-ion exclusion (GCE) model─derived from the solution-friction model─is used to calculate the salt permeability based on the co-ion partitioning coefficients. Our results show that the GCE model predicts the salt permeabilities in RO experiments relatively well, indicating that co-ion partitioning, not salt partitioning, governs salt transport through RO membranes. Our study provides an in-depth understanding of ion partitioning in polyamide RO membranes and its relationship with salt transport.

QCM sensor provides insight into the role of pivotal ions in cellular regulatory volume decrease.

All vertebrate cells generally self-regulate for sustaining homeostasis and cell functions. As a major regulatory mechanism, regulatory volume decrease (RVD) occurs in hypotonicity-induced cell swelling, and then shrinking by the efflux of intracellular osmolytes and water, in which the ions K+, Cl-, and Ca2+ play a key role in the RVD process. We observed that these pivotal ions could result in novel RVD behaviors under repeatedly hypotonic stimulation. However, there is a lack of valid means for assessing the effect of pivotal ions on RVD. In this work, we proposed an effective measurement process based on a quartz crystal microbalance (QCM) combined with cell function of RVD for revealing acute variations in cell volume regulation induced by the pivotal ions. A QCM sensor was implemented by adhering MCF-7 cells to a poly-l-lysine-modified gold chip and cyclic stimulation with hypotonic NaCl medium, in which a frequency shift (Δf) showed the superior feasibility of the technique in exhibiting RVD behaviors. With the increase in the number of cycles, the RVD values decreased progressively under three stimulation cycles with hypotonic NaCl alone. Compared with the first cycle, the RVD level in the second and third cycles declined by 60.7±1.7% and 82.1±1.6% (n=3), respectively; conversely, it recovered in NaCl-KCl solution, but was significantly enhanced by 52.2±0.8% in NaCl-CaCl2 solution. Moreover, the inhibition of chloride channels to block Cl- efflux also decreased the RVD level by 56.2±3.0%. The results indicate that these ions (K+, Cl-, Ca2+) are all able to affect the function of RVD, among which intracellular Cl- depletion reduced RVD during measurement, but which recovered with K+ supplement, and Ca2+ enhanced RVD due to activation of ion channels. Therefore, this work provides a comprehensive assessment of cellular behavior and offers an innovative method for gaining insight into cellular functions and mechanisms. A novel strategy was conducted by integrating a quartz crystal microbalance (QCM) with the function of cell volume regulation for analyzing the role of the pivotal ions ( K+, Cl-, Ca2+) in NaCl media on the behaviors of regulatory cell volume decrease (RVD).

Histidine-Functionalized Diblock Copolymer Nanoparticles Exhibit Enhanced Adsorption onto Planar Stainless Steel.

RAFT aqueous emulsion polymerization of isopropylideneglycerol monomethacrylate (IPGMA) is used to prepare a series of PGEO5MA46 -PIPGMAy nanoparticles, where PGEO5MA is a hydrophilic methacrylic steric stabilizer block bearing pendent cis-diol groups. TEM studies confirm a spherical morphology while dynamic light scattering (DLS) analysis indicated that the z-average particle diameter can be adjusted by varying the target degree of polymerization for the core-forming PIPGMA block. Periodate oxidation is used to convert the cis-diol groups on PGEO5MA46 -PIPGMA500 and PGEO5MA46 -PIPGMA1000 nanoparticles into the analogous aldehyde-functionalized nanoparticles, which are then reacted with histidine via reductive amination. In each case, the extent of functionalization is more than 99% as determined by 1 H NMR spectroscopy. Aqueous electrophoresis studies indicate that such derivatization converts initially neutral nanoparticles into zwitterionic nanoparticles with an isoelectric point at pH 7. DLS studies confirm that such histidine-derivatized nanoparticles remain colloidally stable over a wide pH range. A quartz crystal microbalance is employed at 25°C to assess the adsorption of both the cis-diol- and histidine-functionalized nanoparticles onto planar stainless steel at pH 6. The histidine-bearing nanoparticles adsorb much more strongly than their cis-diol counterparts. For the highest adsorbed amount of 70.5 mg m-2 , SEM indicates a fractional surface coverage of 0.23 for the adsorbed nanoparticles.

A novel molecular imprinted QCM sensor based on MoS2NPs-MWCNT nanocomposite for zearalenone determination.

Zearalenone (ZEN) is a mycotoxin that has a carcinogenic effect and is often found at a high rate in frequently consumed foods. In this study, a characteristic molecular imprinted quartz crystal microbalance (QCM) sensor based on molybdenum disulfide nanoparticle (MoS2NPs)-multiwalled carbon nanotube (MWCNT) nanocomposite (MoS2NPs-MWCNTs) is presented for selective determination of ZEA in rice samples. Firstly, molybdenum disulfide nanoparticle (MoS2NP)-multiwalled carbon nanotube nanocomposites were characterized by using microscopic, spectroscopic, and electrochemical techniques. Then, ZEA-imprinted QCM chip was prepared in the presence of methacryloylamidoglutamicacid (MAGA) as monomer, N,N'-azobisisobutyronitrile (AIBN) as initiator, and ZEA as target molecule by using UV polymerization. The sensor revealed a linearity toward ZEA in the range 1.0-10.0 ng L-1 with a detection limit (LOD) of 0.30 ng L-1. The high repeatability, reusability, selectivity, and stability of the developed sensor enable reliable ZEA detection in rice samples.

Selective Detection of Erythrocytes with QCMs-ABO Blood Group Typing.

Blood transfusion, as well as organ transplantation, is only possible after prior blood group (BG) typing and crossmatching. The most important blood group system is that of Landsteiner's ABO classification based on antigen presence on the erythrocyte surfaces. A mass sensitive QCM (quartz crystal microbalance) sensor for BG typing has been developed by utilizing molecular imprinting technology. Polyvinylpyrrolidone (crosslinked with N,N-methylenebisacrylamide) is a favorable coating that was imprinted with erythrocytes of different blood groups. In total, 10 MHz quartz sheets with two resonators, one for MIP (molecularly imprinted polymer) and the other for NIP (non-imprinted polymer) were fabricated and later used for mass-sensitive measurements. The structure of erythrocyte imprints resembles a donut, as identified by AFM (atomic force microscope). All the erythrocytes of the ABO system were chosen as templates and the responses to these selective coatings were evaluated against all blood groups. Each blood group can be characterized by the pattern of responses in an unambiguous way. The results for blood group O are remarkable given that all types of erythrocytes give nearly the same result. This can be easily understood as blood group O does not possess neither antigen A nor antigen B. The responses can be roughly related to the number of respective antigens on the erythrocyte surface. The imprints generate hollows, which are used for reversible recognition of the erythrocytes. This procedure is based on molecular recognition (based on supramolecular strategies), which results from size, shape and enthalpic interactions between host and guest molecules.

Acetone Detection and Classification as Biomarker of Diabetes Mellitus Using a Quartz Crystal Microbalance Gas Sensor Array.

A gas sensor array was developed and evaluated using four high-frequency quartz crystal microbalance devices (with a 30 MHz resonant frequency in fundamental mode). The QCM devices were coated with ethyl cellulose (EC), polymethylmethacrylate (PMMA), Apiezon L (ApL), and Apiezon T (ApT) sensing films, and deposited by the ultrasonic atomization method. The objective of this research was to propose a non-invasive technique for acetone biomarker detection, which is associated with diabetes mellitus disease. The gas sensor array was exposed to methanol, ethanol, isopropanol, and acetone biomarkers in four different concentrations, corresponding to 1, 5, 10, and 15 µL, at temperature of 22 °C and relative humidity of 20%. These samples were used because human breath contains them and they are used for disease detection. Moreover, the gas sensor responses were analyzed using principal component analysis and discriminant analysis, achieving the classification of the acetone biomarker with a 100% membership percentage when its concentration varies from 327 to 4908 ppm, and its identification from methanol, ethanol, and isopropanol.

Effect of Noise on Determining Ultrathin-Film Parameters from QCM-D Data with the Viscoelastic Model.

Quartz crystal microbalance with dissipation monitoring (QCM-D) is a well-established technique for studying soft films. It can provide gravimetric as well as nongravimetric information about a film, such as its thickness and mechanical properties. The interpretation of sets of overtone-normalized frequency shifts, ∆f/n, and overtone-normalized shifts in half-bandwidth, ΔΓ/n, provided by QCM-D relies on a model that, in general, contains five independent parameters that are needed to describe film thickness and frequency-dependent viscoelastic properties. Here, we examine how noise inherent in experimental data affects the determination of these parameters. There are certain conditions where noise prevents the reliable determination of film thickness and the loss tangent. On the other hand, we show that there are conditions where it is possible to determine all five parameters. We relate these conditions to the mathematical properties of the model in terms of simple conceptual diagrams that can help users understand the model's behavior. Finally, we present new open source software for QCM-D data analysis written in Python, PyQTM.

Unraveling Membrane-Disruptive Properties of Sodium Lauroyl Lactylate and Its Hydrolytic Products: A QCM-D and EIS Study.

Membrane-disrupting lactylates are an important class of surfactant molecules that are esterified adducts of fatty acid and lactic acid and possess industrially attractive properties, such as high antimicrobial potency and hydrophilicity. Compared with antimicrobial lipids such as free fatty acids and monoglycerides, the membrane-disruptive properties of lactylates have been scarcely investigated from a biophysical perspective, and addressing this gap is important to build a molecular-level understanding of how lactylates work. Herein, using the quartz crystal microbalance-dissipation (QCM-D) and electrochemical impedance spectroscopy (EIS) techniques, we investigated the real-time, membrane-disruptive interactions between sodium lauroyl lactylate (SLL)-a promising lactylate with a 12-carbon-long, saturated hydrocarbon chain-and supported lipid bilayer (SLB) and tethered bilayer lipid membrane (tBLM) platforms. For comparison, hydrolytic products of SLL that may be generated in biological environments, i.e., lauric acid (LA) and lactic acid (LacA), were also tested individually and as a mixture, along with a structurally related surfactant (sodium dodecyl sulfate, SDS). While SLL, LA, and SDS all had equivalent chain properties and critical micelle concentration (CMC) values, our findings reveal that SLL exhibits distinct membrane-disruptive properties that lie in between the rapid, complete solubilizing activity of SDS and the more modest disruptive properties of LA. Interestingly, the hydrolytic products of SLL, i.e., the LA + LacA mixture, induced a greater degree of transient, reversible membrane morphological changes but ultimately less permanent membrane disruption than SLL. These molecular-level insights support that careful tuning of antimicrobial lipid headgroup properties can modulate the spectrum of membrane-disruptive interactions, offering a pathway to design surfactants with tailored biodegradation profiles and reinforcing that SLL has attractive biophysical merits as a membrane-disrupting antimicrobial drug candidate.

Quartz Crystal Microbalance Platform for SARS-CoV-2 Immuno-Diagnostics.

Rapid and accurate serological analysis of SARS-CoV-2 antibodies is important for assessing immune protection from vaccination or infection of individuals and for projecting virus spread within a population. The quartz crystal microbalance (QCM) is a label-free flow-based sensor platform that offers an opportunity to detect the binding of a fluid-phase ligand to an immobilized target molecule in real time. A QCM-based assay was developed for the detection of SARS-CoV-2 antibody binding and evaluated for assay reproducibility. The assay was cross-compared to the Roche electrochemiluminescence assay (ECLIA) Elecsys® Anti-SARS-CoV-2 serology test kit and YHLO's chemiluminescence immunoassay (CLIA). The day-to-day reproducibility of the assay had a correlation of r2 = 0.99, p < 0.001. The assay linearity was r2 = 0.96, p < 0.001, for dilution in both serum and buffer. In the cross-comparison analysis of 119 human serum samples, 59 were positive in the Roche, 52 in the YHLO, and 48 in the QCM immunoassay. Despite differences in the detection method and antigen used for antibody capture, there was good coherence between the assays, 80-100% for positive and 96-100% for negative test results. In summation, the QCM-based SARS-CoV-2 IgG immunoassay showed high reproducibility and linearity, along with good coherence with the ELISA-based assays. Still, factors including antibody titer and antigen-binding affinity may differentially affect the various assays' responses.

Lipid Membrane Remodeling by the Micellar Aggregation of Long-Chain Unsaturated Fatty Acids for Sustainable Antimicrobial Strategies.

Antimicrobial fatty acids derived from natural sources and renewable feedstocks are promising surface-active substances with a wide range of applications. Their ability to target bacterial membrane in multiple mechanisms offers a promising antimicrobial approach for combating bacterial infections and preventing the development of drug-resistant strains, and it provides a sustainable strategy that aligns with growing environmental awareness compared to their synthetic counterparts. However, the interaction and destabilization of bacterial cell membranes by these amphiphilic compounds are not yet fully understood. Here, we investigated the concentration-dependent and time-dependent membrane interaction between long-chain unsaturated fatty acids-linolenic acid (LNA, C18:3), linoleic (LLA, C18:2), and oleic acid (OA, C18:1)-and the supported lipid bilayers (SLBs) using quartz crystal microbalance-dissipation (QCM-D) and fluorescence microscopy. We first determined the critical micelle concentration (CMC) of each compound using a fluorescence spectrophotometer and monitored the membrane interaction in real time following fatty acid treatment, whereby all micellar fatty acids elicited membrane-active behavior primarily above their respective CMC values. Specifically, LNA and LLA, which have higher degrees of unsaturation and CMC values of 160 µM and 60 µM, respectively, caused significant changes in the membrane with net |Δf| shifts of 23.2 ± 0.8 Hz and 21.4 ± 0.6 Hz and ΔD shifts of 5.2 ± 0.5 × 10-6 and 7.4 ± 0.5 × 10-6. On the other hand, OA, with the lowest unsaturation degree and CMC value of 20 µM, produced relatively less membrane change with a net |Δf| shift of 14.6 ± 2.2 Hz and ΔD shift of 8.8 ± 0.2 × 10-6. Both LNA and LLA required higher concentrations than OA to initiate membrane remodeling as their CMC values increased with the degree of unsaturation. Upon incubating with fluorescence-labeled model membranes, the fatty acids induced tubular morphological changes at concentrations above CMC. Taken together, our findings highlight the critical role of self-aggregation properties and the degree of unsaturated bonds in unsaturated long-chain fatty acids upon modulating membrane destabilization, suggesting potential applications in developing sustainable and effective antimicrobial strategies.