Ratio Of Beads Research Articles

B-025 Revolutionizing NGS-driven Laboratory Developed Tests: The impact of automated liquid handling on plate-based bead clean-up

Abstract Background Automated liquid handling systems are proving to be a transformative technology in the landscape of Laboratory Developed Tests (LDTs), especially those utilizing Next-Generation Sequencing (NGS). By enhancing accuracy, boosting efficiency, and offering scalable solutions, these systems enable labs to meet current and future challenges more effectively. The right system not only improves the quality of diagnostics but also positions labs to be agile and adaptive in a rapidly advancing healthcare environment. firefly® is a well-suited liquid handling solution for diagnostics labs developing LDTs due to its compact footprint, user-friendly software and exceptional versatility. Validated protocol templates that support NGS library preparation are readily available from the firefly cloud and can be adapted to meet specific lab needs. For example, automated bead-based clean-ups designed for 96- and 384- NGS applications are available on firefly. These remove DNA fragments of undesirable sizes, enzymes or primers rom the amplified DNA after a PCR reaction. Automating this process facilitates higher throughput and scaling. firefly has a unique combination of 6 positive displacement dispense heads and a 96/384-well air-displacement pipetting head, making it ideal to distribute the reagents required for bead clean-ups. Methods firefly protocols were developed and optimized to be compatible with a range of elution volumes and bead ratios. Protocol performance was tested using library DNA (∼300bp). The same input DNA was plated out and quantified using the QuantiFluor dsDNA system (Promega) before and after running the protocol. This data was used to assess any reduction in DNA concentration and uniformity across the plate. The protocol's ability to successfully modulate the fragment size distribution was demonstrated using a 50bp DNA ladder and different bead ratios. Data was obtained using a Fragment Analyzer (Agilent, DNF-474 HS NGS Fragment Kit). Well-to-well contamination was assessed by running no-template-controls (NTCs) in wells adjacent to library DNA, then analyzed by qPCR using a LightCycler 480 System (Roche, KAPA Library Quantification kit). Results 96 format has a setup time of 10 minutes, run time of 43 and %CV library concentration of <6%. 384 format has a setup time of 10 minutes, run time of 47 and %CV library concentration of <8%. Uniformity of concentration and fragment sizes was seen across the plates. Comparison of the library concentration before and after the clean-up showed a 5% reduction in DNA concentration after the 96-well plate clean-up and an 11% reduction in library concentration when using the 384-well plate clean-up. Fragment size analysis showed that fragment sizes can be successfully modulated on firefly by changing the bead ratio of the firefly run and no detectable well-to-well contamination was detected by qPCR. Conclusions We have demonstrated that automated bead clean-ups on firefly are uniform show uniformity across both 96- and 384-well plates, with no detectable well-to-well contamination, to deliver the consistent and robust results required for an LDT. Bead-based cleanups on firefly offer a fast, 100% walkaway solution. By running 96-samples at once on firefly, we see approximately a 6-fold increase in throughput than when performed manually by a single user.

Core-shell structured alginate-based hydrogel beads modified by starch and protocatechuic acid: Preparation, characterization, phenolic slow release and stable antioxidant potential

A novel core-shell structured alginate-based hydrogel bead modified by co-gelatinizing with starch and protocatechuic acid (PA), was designed to modulate physical properties of beads, release behavior and antioxidant stability of encapsulated bioactives. Core was fabricated by ionotropic gelation, and its formulation (ratio of sodium alginate/starch) was determined by particle size/starch distribution, texture and bioactive encapsulation capacity of core. Then, coating core with shell-forming solution co-gelatinized with different doses of PA, and subsequently cross-linked with Ca2+ to obtain core-shell structured beads. Surface microstructure, mechanical characteristics, and swelling ratio of beads were affected by concentrations of PA. Besides, core-shell structure containing PA could enhance delivery and sustained release of encapsulated phenolic bioactives during in vitro digestion, and improve their antioxidant potential stability. Furthermore, interaction between PA and polysaccharide components was elucidated by FTIR and TGA. The present information was beneficial for the advancement of functional food materials and bioactive delivery systems.

Preparation and Characterization of Silica-Coated Sodium Alginate Hydrogel Beads and the Delivery of Curcumin

In this study, to address the defects of sodium alginate (SA), such as its susceptibility to disintegration, silica was coated on the outer layer of sodium alginate hydrogel beads in order to improve its swelling and slow-release properties. Tetraethyl orthosilicate (TEOS) was used as the hydrolyzed precursor, and the solution of silica precursor was prepared by sol-gel reaction under acidic conditions. Then SA–silica hydrogel beads prepared by ionic crosslinking method were immersed into the SiO2 precursor solution to prepare SA–silica hydrogel beads. The chemical structure and morphology of the hydrogel beads were characterized by XRD, FTIR, and SEM, and the results showed that the surface of SA–silica beads was successfully encapsulated with the outer layer of SiO2, and the surface was smooth and dense. The swelling experiments showed that the swelling performance effectively decreased with the increase of TEOS molar concentration, and the maximum swelling ratio of the hydrogel beads decreased from 41.07 to 14.3, and the time to reach the maximum swelling ratio was prolonged from 4 h to 8 h. The sustained-release experiments showed that the SA–silica hydrogel beads possessed a good pH sensitivity, and the time of sustained-release was significantly prolonged in vitro. Hemolysis and cytotoxicity experiments showed that the SA-silica hydrogel beads were biocompatible when the TEOS molar concentration was lower than 0.375 M. The SA–silica-2 hydrogel beads had good biocompatibility, swelling properties, and slow-release properties at the same time.

Efficient recovery of beta-mannanase fused with LysM3014 domain in Escherichia coliby glass bead based-cell disruption method

Toward the development of whole cell biocatalysts via non-GMO strategy, in this study, three cell disruption methods (sonication, freezingand thawing and bead-shaking) have been evaluatedto recover efficiently fusion protein LysM3014-ManB containing a beta-mannanase (ManB) from Bacillus lichenifomisfused with a single LysM3014 domain derived from a putative extracellulartransglycosylase Lp_3014 of Lactobacillus plantarumWCFS1. The results showed that the highest volumetric activity of fusion protein (~46651 U/lfermentation) was obtained when shaking the recombinant E. coliwith glass bead. Moreover, the enzymatic activity of LysM3014-ManB increased approximately 1.4 folds(~66000 U/lfermentation)under the optimum conditions of glass bead-based cell disruption (falcon 50 ml, glass bead ratio of 1.0 g/mland disruption time of 8 min). The study indicates that shaking the cells with glass bead is likely to be the simplest and the most efficient cell disruption methodin comparison with other disruption methods.

Controlling the Porosity of Starch Hydrogels with Poly(Methyl Methacrylate) (PMMA) Beads

AbstractStarch is a natural, biodegradable polymer that can be used to prepare hydrogels with various applications in food, pharmacy, medicine, agriculture, etc. In this study, a method of preparation using poly(methyl methacrylate) (PMMA) beads to control the porosity of starch hydrogels is proposed. The hydrogels are crosslinked with trisodium trimetaphosphate and dried in a vacuum oven. Results show that increasing the amount of PMMA beads result in higher porosity hydrogels ranging from ≈35% for hydrogels where no PMMA beads are used to ≈88% for hydrogels where the mass ratio of PMMA beads to starch is 10:1. Higher porosity hydrogels have a higher equilibrium water content and swelling degree, but lower mechanical properties. All hydrogels have a low solubility (<≈5%) and a high gel fraction (>≈90%) percentage. Upon degradation in α‐amylase at 37 °C, low porosity hydrogels (prepare with 0:1 and 1:10 PMMA beads:starch) degrade within 30 min, while high porosity hydrogels (prepare with 1:1 and 10:1 PMMA beads:starch) degrade within 3 weeks. The release of a dye that is incorporated into the hydrogel walls follows similar kinetics. Therefore, the use of PMMA beads is an efficient method to control starch hydrogel's porosity and properties.

Establishing and verifying the threshold value of HLA mixed antigen reagent screening test results

Objective: To establish the threshold value of human leukocyte antigen (HLA) mixed antigen reagent screening test results, and to verify it by HLA single antigen reagent confirmation test results. Methods: The results of 2 255 serum samples tested for HLA antibodies by HLA mixed antigen reagent in the department of HLA Laboratory, the First Affiliated Hospital of Soochow University from October 2017 to December 2021 were retrospectively analyzed. Among them, 1 139 samples were also tested by single antigen HLA Class-Ⅰ reagent and 1 116 samples were also tested by single antigen HLA Class-Ⅱ reagent. Based on the same antigens coated with both reagents, the Mean Fluorescence Intensity (MFI) and Nomalized Background ratio (NBG ratio) of 12 HLA Class-Ⅰ beads and 5 HLA Class-Ⅱ beads in the HLA mixed antigen reagent and the MFI of 77 anti-HLA class-Ⅰ antibodies and 35 anti-HLA class-Ⅱ antibodies detected by HLA single antigen reagent were recorded. The MFI and NBG ratio of HLA mixed antigen reagent beads in 1 139 or 1 116 samples were segmented according to the positive rate of antibodyies detected by the single antigen reagent corresponding to the antigens coated with each HLA mixed antigen reagent bead, and the results of the HLA mixed antigen screening test were verified by the HLA single antigen reagent confirmation test. Results: The threshold values of MFI and NBG ratio of HLA mixed antigen reagent's 17 beads were established. The MFI of No. 1 to No. 17 beads of HLA mixed antigen reagent ranged from 26.86 to 21 925.58, and the NBG ratio ranged from 0 to 434.65. According to the positive detection rate of HLA single antigen reagent corresponding to the coated antigens, the MFI and NBG ratio of the beads of HLA mixed antigen reagent were divided into positive interval, suspicious positive interval, suspicious negative interval and negative interval. The positive rates of anti-HLA class-Ⅰ antibodies by HLA mixed antigen reagent and single antigen HLA Class-Ⅰ reagent were 87.5% (997/1 139) and 66.3% (755/1 139). The positive rates of anti-HLA class-Ⅱ antibodies were 63.4% (707/1 116) and 44.9% (501/1 116). In the samples with suspicious negative, suspicious positive and positive results of HLA class-Ⅰ、Ⅱ antibodies detected by HLA mixed antigen reagent, the positive detection rates of single antigen HLA Class-Ⅰ reagent were 14.9% (17/114), 41.3% (145/351) and 91.3% (590/646), respectively. The positive detection rates of single antigen HLA Class-Ⅱ reagent were 15.5% (58/375), 26.5% (81/306) and 88.8% (356/401), respectively. Conclusions: In this study, the threshold values of MFI and NBG ratio of HLA mixed antigen reagent screening test are established, and the threshold values are verified by the results of HLA single antigen reagent confirmation test. HLA mixed reagent screening test can be used for screening of HLA antibodies, and if necessary, it should be combined with HLA single antigen confirmatory test for clinical detection of HLA antibodies.

Unlocking new dimensions in long-acting injectables using lipid mesophase-based beads

In this study, we explored the use of lipid mesophases (LMPs) as a biocompatible and biodegradable material for sustained drug delivery. Our hypothesis centered on leveraging the high surface-to-volume ratio of LMP-based beads to enhance strength, stability, and surface interaction compared to the LMP bulk gel. To modulate drug release, we introduced antioxidant vitamin E into the beads, influencing mesophase topologies and controlling drug diffusion coefficients. Four drugs with distinct chemical properties and intended for three different pathologies and administration routes were successfully loaded into the beads with a drug entrapment efficiency exceeding 80 %. Notably, our findings revealed sustained drug release, irrespective of the drugs' chemical properties, culminating in the development of an injectable formulation. This formulation allows direct administration into the target site, minimizing systemic exposure, and thereby mitigating adverse effects. Our approach demonstrates the potential of LMP-based beads for tailored drug delivery systems with broad applications in diverse therapeutic scenarios.

Limits of the detection of microplastics in fish tissue using stimulated Raman scattering microscopy.

We demonstrate the detection sensitivity of microplastic beads within fish tissue using stimulated Raman scattering (SRS) microscopy. The intrinsically provided chemical contrast distinguishes different types of plastic compounds within fish tissue. We study the size-dependent signal-to-noise ratio of the microplastic beads and determine a lower boundary for the detectable size. Our findings demonstrate how SRS microscopy can serve as a complementary modality to conventional Raman scattering imaging in order to detect and identify microplastic particles in fish tissue.

Shotgun Characterization of the Circulating IgM Antigenome of an Infectious Pathogen by Immunocapture-LC-MS/MS from Dried Serum Spots.

One of the main challenges in compiling the complete collection of protein antigens from pathogens for the selection of vaccine candidates or intervention targets is to acquire a broad enough representation of them to be recognized by the highly diversified immunoglobulin repertoire in human populations. Dried serum spot sampling (DSS) retains a large repertoire of circulating immunoglobulins from each individual that can be representative of a population, according to the sample size. In this work, shotgun proteomics of an infectious pathogen based on DSS sampling coupled with IgM immunoprecipitation, liquid chromatography-mass spectrometry (LC-MS/MS), and bioinformatic analyses was combined to characterize the circulating IgM antigenome. Serum samples from a malaria endemic region at different clinical statuses were studied to optimize IgM binding efficiency and antibody leaching by varying serum/immunomagnetic bead ratios and elution conditions. The method was validated using Plasmodium falciparum extracts identifying 110 of its IgM-reactive antigens while minimizing the presence of human proteins and antibodies. Furthermore, the IgM antigen recognition profile differentiated between malaria-infected and noninfected individuals at the time of sampling. We conclude that a shotgun proteomics approach offers advantages in providing a high-throughput, reliable, and clean way to identify IgM-recognized antigens from trace amounts of serum. The mass spectrometry raw data and metadata have been deposited with ProteomeXchange via MassIVE with the PXD identifier PXD043800.

In silico study of area fraction effects on the behaviour of a ferrogranulate layer in an orthogonal applied field

In our study, we explore ferrogranulate—a mix of glass and steel particles, specifically Stockmayer and repulsive spheres. Confined to a monolayer with three-dimensional rotation, magnetic particles respond to an external magnetic field applied orthogonal to the granulate plane. By varying the overall area fraction while maintaining a constant steel-to-glass bead ratio, we analyze cluster formation. Results show that concentration growth both slows cluster dynamics and increases the likelihood of larger clusters, hindering the response to the magnetic field. At low applied field values, our calculations suggest a bistable region at high concentrations, where droplet precursors form due to dipolar and steric interactions.

Immuno-capture of biomolecules on functionalized magnetic beads: From characterization to fluorescent detection of a biomarker for Alzheimer's disease toward high performance biosensing

It is reported herein a new approach to study the orientation and density of mouse antibody grafting on magnetic beads, serving for immunoassays and biosensors with fluorescent detection of biomolecules. This approach is based on selective enzymatic digestion of target grafted antibodies at a specific site below the hinge position to provide F(ab′)2 and Fc fragments, followed by separation and determination of these fragments with size exclusion chromatography (SEC) coupled with fluorescence detection (FLD). The developed method was applied for evaluation of immunoglobulin (IgG2a) grafting capacity on three different biofunctionalized magnetic beads (i.e., Tosyl-activated, carboxylic, protein G). Tosyl-activated and protein G beads at different optimal grafting IgG: bead ratios (i.e., 110 µg: 1000 µg and 240 µg: 1000 µg, respectively) exhibited superior grafting capacity than carboxylic counterparts. Under the optimized conditions, more than 70 % of antibodies were grafted on tosyl-activated and protein G beads in the right orientation. This approach was then demonstrated with different commercially available antibodies specific to amyloid-beta peptide 1–42 (Aβ1–42) for magneto-immunoassays and fluorescent detection of this peptide that is an established biomarker for molecular diagnosis of Alzheimer's disease.

A Modified Method for Transient Transformation via Pollen Magnetofection in Lilium Germplasm.

Lily (Lilium spp.) is a popular ornamental plant. Traditional genetic transformation methods have low efficiency in lily, thus development of a high-efficiency genetic transformation system is important. In this study, a novel transient transformation method involving pollen magnetofection was established and optimized pollen viability, and exogenous gene expression in magnetofected pollen and that of different germplasm were assessed. The highest germination percentage of Lilium regale pollen was 85.73% in medium containing 100 g/L sucrose, 61.5 mg/L H3BO3, and 91.5 mg/L CaCl2. A 1:4 ratio of nanomagnetic beads to DNA plasmid and transformation time of 0.5 h realized the highest transformation efficiency (88.32%). The GFP activity in transformed pollen averaged 69.66%, while that of the control pollen was 0.00%. In contrast to the control, transgenic seedlings obtained by pollination with magnetofected pollen showed strong positive GUS activity with 56.34% transformation efficiency. Among the lily germplasm tested, 'Sweet Surrender' and L. leucanthum had the highest transformation efficiency (85.80% and 54.47%), whereas L. davidii var. willmottiae was not successfully transformed. Transformation efficiency was positively correlated with pollen equatorial diameter and negatively correlated with polar axis/equatorial diameter ratio. The results suggest that pollen magnetofection-mediated transformation can be applied in Lilium but might have species or cultivar specificity.

Determination of four aflatoxins in feeds by high throughput automated immunoaffinity magnetic beads purification-ultra performance liquid chromatography

Aflatoxin (AFT) is an extremely toxic and highly toxic carcinogenic substance. This is particularly problematic due to the risk of aflatoxin contamination in raw feed materials and products during production, transportation, and storage. In this study, immunoaffinity magnetic beads (IMBs) were prepared for the purification of four aflatoxins (aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1) and aflatoxin G2 (AFG2)). The aflatoxin contents were then determined rapidly and accurately using ultra performance liquid chromatography (UPLC). More specifically, the coupling ratio of magnetic beads (MBs) to the aflatoxin monoclonal antibody was initially optimized, wherein an MB volume of 1 mL and an antibody content of 2.0 mg was found to meet the purification requirements of this method. The magnetic properties of the MBs and the IMBs were then investigated using a vibrating sample magnetometer (VSM) at room temperature. As a result, the maximum saturation super magnetizations of the MBs and the IMBs were determined to be 28.61 and 23.22 emu/g, respectively, indicating that the saturation magnetization intensity of the IMBs was reduced by coupling with a non-magnetic antibody. However, the saturation magnetization intensity remained sufficiently high to permit magnetic separation from the solution. In addition, the appearance of the IMBs was examined using a biomicroscope, and it was clear that the magnetic cores were wrapped in agarose gel. Furthermore, the reaction time between the IMBs and the aflatoxins was investigated, and the optimal reaction time for meeting the purification requirements was determined to be 2 min. The stability of the IMBs was then evaluated under refrigerated storage conditions at 4 ℃. It was found that the prepared IMBs maintained a high aflatoxin enrichment capacity for at least eight months. Through the examination of three different extraction solutions, a mixture of acetonitrile and water (70∶30, v/v) was found to be optimal for the extraction of aflatoxins from the feed samples. Moreover, five sample dilutions and purification effects were also examined, and phosphate-buffered saline (containing 0.5% Tween-20) was selected as the preferred sample dilutant. With the optimized conditions, the effectiveness of using IMB for the purification of different feed samples was investigated. The resulting UPLC chromatogram showed no spurious peaks close to the target peaks, demonstrating a good purification performance. Following matrix spiking (5, 20, and 40 μg/kg, calculated based on AFB1) of the four feed samples (i. e., soybean meal, distillers dried grains with solubles, pig feed, and chicken feed), the spiked recoveries of the four aflatoxins ranged from 91.1% to 119.4% with a relative standard deviation (RSD) of <6.9%. In addition, the inter-day precision was 4.5% to 7.5%, and the method exhibited a good reproducibility. Subsequently, the developed method was used to detect AFB1 using reference materials. The test value was 18.6 μg/kg with an accuracy of 110.3%, thereby constituting satisfactory results. Upon testing 21 randomly purchased feed samples using this method, four of these samples contained AFB1, and the test results obtained using the developed method and stable isotope dilution LC-MS/MS were comparable. It was therefore apparent that the IMB purification method combined with UPLC analysis exhibited a good accuracy for aflatoxin determination. Thus, an automatic purification system was established to facilitate the operation and use of IMBs. This system was able to purify 24 samples simultaneously in 30 min. An IMB purification kit for was also designed and produced for aflatoxin detection in feed samples. The kit contained the sample dilutant, IMBs, the washing solution, and the eluent. After extraction of the feed sample, the extraction solution was added to the sample wells provided in the kit, and the purification system automatically completed the steps of aflatoxin enrichment, impurity washing, and elution of the target toxin. It should be noted that the purification process does not require the operator to manually add the solution, thereby simplifying operation. Overall, the purification method established in this study achieved the high-throughput and automatic purification of the four aflatoxins in feed samples.

Development of an AlphaLISA assay for sensitive and accurate detection of influenza B virus.

Influenza B virus (IBV) is highly contagious, spreads rapidly, and causes seasonal epidemic respiratory disease in the human population, especially in immunocompromised people and young children. Clinical manifestations in this high-risk population are often more severe than in immunocompetent hosts and sometimes atypical. Therefore, rapid, and accurate detection of IBV is important. An amplified luminescent proximity homogeneous assay linked immunosorbent assay (AlphaLISA) was developed for detection of IBV by optimizing the ratio of IBV antibody-labeled receptor beads, streptavidin-conjugated donor beads and biotinylated IBV antibody, as well as the optimal temperature and time conditions for incubation. Assay sensitivity, specificity and reproducibility were evaluated. A total of 228 throat swab samples and inactivated influenza B virus were tested by AlphaLISA and lateral flow colloidal gold-based immunoassay (LFIA). AlphaLISA produced the best results for detection of inactivated influenza B virus when IBV antibody-labeled acceptor beads were 50 μg/ mL, streptavidin-conjugated donor beads were 40 μg/mL, and biotinylated IBV antibody was 0.5 μg/mL at 37°C for 15-10 min. Under these conditions, AlphaLISA had a limit of detection of 0.24 ng/mL for the detection of influenza B nucleoprotein, did not cross react with other common respiratory viruses, and showed good reproducibility with inter-assay coefficient of variation (CV) and intra-assay CV < 5%. The results of 228 clinical throat swab samples showed good agreement between AlphaLISA and LFIA (Kappa = 0.982), and AlphaLISA showed better sensitivity than LFIA for detecting inactivated influenza B virus. AlphaLISA showed higher sensitivity and throughput in the detection of IBV and can be used for IBV diagnosis and epidemic control.

Design of Spherical Gel-Based Magnetic Composites: Synthesis and Characterization

The purpose of the study was the synthesis and the physicochemical characterization of spherical beads of magnetically active composite ferrogels (FGs) with diameters of 2–3 mm for further application to the needs of targeted drug delivery and/or replacement therapy. Spherical FGs based on a physical network of calcium alginate (CaAlg), a chemical network of polyacrylamide (PAAm), and a combined network of calcium alginate and polyacrylamide (PAAm/CaAlg) were analyzed. FGs were filled with γ-Fe2O3 magnetic nanoparticles (MNPs) obtained by using the electrical explosion of wire method. A comparative study of the swelling behavior and of the structural features of the polymeric network in CaAlg, PAAm/CaAlg, and PAAm spherical beads was performed. It was shown that the densest network was provided by a combination of chemical and physical networking in PAAm/CaAlg FGs. If the physical network were removed from FGs it resulted in a substantial increase in the average diameter and the swelling ratio of spherical beads and a decrease in the MNPs concentration in the swollen FGs by approximately two times. It was shown that irrespective of the gel composition, the embedding of maghemite nanoparticles led to an increase in the swelling ratio of the polymeric network. This indicated the absence of strong intermolecular interactions between the polymer and the filler. The results obtained might be useful for the design of magnetically active spherical FG beads of a given size and controlled physicochemical properties.

Kinetics of formation of a macroscale binary Coulombic material

This paper describes the formation kinetics of a two-dimensional, binary Coulombic material. The material is formed by mechanically agitating millimeter-sized nylon and polytetrafluoroethylene (PTFE) beads that tribocharge positively and negatively, respectively. The authors alter the relative number of nylon and PTFE beads, without changing their combined total. They discover a common transient structure that does not depend on the relative ratio of nylon and PTFE beads, reveal a structure transition driven by the minimization of Coulombic energy, and provide insights for the rational design of materials.

FABRICATION AND CONTROLLING MORPHOLOGY OF POLYETHYLENE OXIDE/SODIUM ALGINATE BEADS AND/OR FIBERS: EFFECT OF VISCOSITY AND CONDUCTIVITY IN ELECTROSPINNING

Objective: The aim of the study is to determine the viscosity and conductivity of PEO/NaAlg polymer solutions, to formulate PEO/NaAlg beads, bead-on-string fibers and fibers via electrospinning, and to perform advanced morphological characterization studies on them.&#x0D; Material and Method: Effect of PEO and NaAlg concentration and ratio of them on spinnability, viscosity and conductivity of solutions, and also on the morphological properties of PEO/NaAlg electrospun fibers, and beads (length, width and aspect ratio of beads, number of beads and bead area) were investigated.&#x0D; Result and Discussion: As a result, electrospun materials were produced using PEO/NaAlg beads and/or fibers, which are valuable for medical and biological applications such as tissue engineering, wound dressing, drug delivery system. Viscosity and conductivity of solutions, and morphological properties of the obtained materials were found to be affected by PEO and NaAlg concentration and the ratio of them. Spinnability was improved thanks to the increase in conductivity in the presence of PEO. The lower viscosity and conductivity resulted in the production of beads that were generally smaller and greater in number in the material and having higher area. The morphological properties of PEO/NaAlg electrospun materials can be modified by controlling the parameters examined in the study.

Synthesis of Silicon Oxycarbide Beads from Alkoxysilane as Anode Materials for Lithium-Ion Batteries.

Silicon is an important anode material for lithium-ion batteries because of its high theoretical capacity. However, the large volume expansion of silicon anodes hinders its commercial utilization. As an alternative, silicon oxycarbides (SiOCs) mitigate the expansion of anodes during lithiation, and the synthesis of SiOC beads from silanes is rather simple and at a low cost. In this study, we compared three different reactor setups for making the SiOC beads from methyltrimethoxysilane (MTMS) and found that the control of residence time was crucial. Thereby, the batch reactor turned out to be the easiest one for making monodispersed beads. We also reduced the O/Si ratio of the SiOC beads by adding dimethyldimethoxysilane (DMDMS) for better battery performance. The first-cycle delithiation capacity of the most stable material was over 1796 mA h/g, with an initial Coulombic efficiency of 82%, while the capacity retention after 170 cycles was 67% (992 mA h/g) at a charging rate of 2 A/g in the potential range of 0.01-3 V. This was among the best of the reported data so far for the SiOC beads from MTMS.

Establishment and application of quantitative immunoassay method based on stable element labeling and inductively coupled plasma mass spectrometry

Objective: To establish a quantitative immunoassay method based on stable element labeling and inductively coupled plasma mass spectrometry (ICP-MS) for the detection of serum amyloid A (SAA) and evaluate its performance. Methods: An immunoassay system based on sandwich method was established with magnetic bead as carrier and holmium (Ho) as element tags. The binding ratio of hydrophilic streptavidin magnetic beads and biotinylated antibody, the amount of elemental antibody, and the reaction time were optimized to choose the optimal reaction conditions. According to the documents of Clinical and Laboratory Standards Institute (CLSI), the analytical performance was evaluated, including the limit of blank (LOB), linearity, accuracy, specificity, imprecision and interference test. Finally, 82 SAA plasma samples were collected after the turbidimetric inhibition immunoassay, and the newly established method was used for detection. Moreover, the detection results of the two methods were analyzed by Pearson correlation analysis. Results: The optimal binding ratio of hydrophilic streptavidin and biotinylated antibody was 1∶0.15, the amount of Ho-labeled antibody was 3 μl and the incubation time of the two reaction steps was 40 min and 30 min, respectively. The LOB was 0.6 ng/ml. The linearity was good within the range of 0-1 200 μg/L (R2=0.998 9, P<0.001). The inter-batch precision of high-value samples and low-value samples was 9.42% and 7.95%, respectively, and the intra-batch precision was 14.56% and 13.56%, respectively. The recovery was 96.01%-104.76%. The cross-reaction rates with procalcitonin (PCT) and C-reactive protein (CRP) were 0.45% and 0.015%, respectively. When the concentration of triglyceride≤35.5 mg/L, bilirubin≤0.52 mg/L and hemoglobin≤2.4 g/L, the interference bias was less than 10%. The results of 82 SAA plasma samples were 12.65 (4.45, 59.03) mg/L by ICP-MS immunoassay and 18.23 (9.33, 68.72) mg/L by turbidimetric inhibition immunoassay, respectively. The newly established system was well correlated with turbidimetric inhibition immunoassay (R2=0.983, P<0.001). Conclusion: The quantitative immunoassay for SAA with Ho as marker established in this study has high precision, good accuracy, high specificity, and wide linear range, which can meet the clinical testing requirements.

Optimizing the Use of Solid-Phase Reversible Immobilization Beads for High-Throughput Full-Length 16S rDNA Sequencing Library Construction

Objective: Solid-phase reversible immobilization (SPRI) beads are widely used for high-throughput sequencing library construction to purify and recover nucleic acids. This research was aimed at investigating the effects of SPRI bead ratio, incubation time, and elution time on nucleic acid recovery during full-length 16S rDNA high-throughput sequencing library construction. Methods: The effects of different SPRI bead ratios, incubation times, and elution times were compared for three different initial sample amounts. An L9(33) orthogonal experiment was designed to determine the optimal combination of these factors. Results: The incubation time of three factors including SPRI beads ratio, incubation time, and elution time had a statistically significant effect on the recovery rate for the initial sample amount of 1500 ng and 3000 ng. The orthogonal experiment results indicated that incubation time had the greatest impact among the three factors. Conclusion: Incubation time significantly influences recovery rate in full-length 16S rDNA high-throughput sequencing library construction. The use of 0.8× SPRI beads, 15 minutes of incubation, and 10 minutes of elution resulted in the highest recovery rate. SPRI beads offer a viable method for recovering full-length 16S rDNA amplicons.