Efficient Elution Research Articles

Ultra-highly efficient enrichment of uranium from seawater via studtite nanodots growth-elution cycle

Consecutive uranium extraction from seawater is a promising approach to secure the long-term supply of uranium and the sustainability of nuclear energy. Here, we report an ultra-highly efficient strategy via studtite nanodots growth with impressive uranyl uptake capacity of ~ 154.50 mg/g from natural seawater in 12 consecutive days (i.e., average for ~ 12.875 mg/g/day). Uranyl can be extracted as studtite under visible light via the reaction between the adsorbed uranyl and the photogenerated H2O2 with imine-based Covalent-Organic Framework photocatalysts. In detail, over Tp-Bpy, Tp-Bpy-2 and Tp-Py with multiple uranyl chelating sites, uranyl is found extracted as studtite nanodots which can be eluted readily, while over Tp-Bd and Tb-Bpy, uranyl is transformed into studtite nanorods that is more inert for elution. Abundant chelating sites of uranyl via structural regulation of COF photocatalysts are proved to facilitate the formation and efficient elution of studtite nanodots.

Optimization and Evaluation of Column Efficiency in RP-HPLC Method for Pharmaceutical and Forensic Drug Analysis

Reverse Phase High Performance Liquid Chromatography (RP-HPLC) is an efficient and reliable technique employed in pharmaceutical and forensic analysis. The assay determination in pharmaceutical and certain molecular detection is performed using this technique. Performance of chromatographic columns play a central role in development of more efficient separation methods. It is one of the main component of system suitability testing in a chromatographic system. The column efficiency can be assessed by using various traditional methods and reproducible results can be evaluated using liquid chromatography We studied the column efficiency using RP-HPLC technique for pharmaceutical drug product Aceclofenac in a validated method. Two mobile phases combinations Acetonitrile: Methanol (80:20 v/v) and Acetonitrile: Methanol: Ammonia (225:50:1 v/v) were compared on C-8 Lichrospher RP-HPLC column for Aceclofenac and three tablet samples (30 ppm). The height equivalent to theoretical plate (HETP) referred as direct measure of column efficiency produced efficient results (530-545 and 190-230) for respective mobile phases. The lower HEPT is an indication of efficient elution in second mobile phase. The change in polarity as per pKa of the analyte may enhance elution and resolution. helpful in method development for pharmaceutical and forensic drug analysis using liquid chromatography.

Polyvinyl Alcohol‐Conjugated L‐Cysteine: A Novel Metal Pincer for Efficient Heavy Metal Ions Removal from Wastewater

AbstractA facilely prepared composite based on Polyvinyl alcohol anchored‐ L‐cysteine (PVA‐L‐CYS) demonstrated efficient functionality in the elimination of heavy metals (Hg, Cr, Pb, and Cd) from working wastewater. The synthesis of the composite was validated through a comprehensive characterization utilizing various analytical techniques to ensure the composite‘s structural, chemical, and physical properties. At ambient temperature, pH of 4–5.5, interaction time of 15–20 minutes, and concentration of 200 mg/L metal ions, the composite exhibited a maximum retention capacity of 48.5, 25, 45.25, and 44.25 mg/g for Hg, Cr, Pb, and Cd, respectively. Langmuir modeling was establish to be more fitting to the practical results than Freundlich, providing theoretical values of 49.02, 25.97, 46.08, and 44.84 mg/g for Hg, Cr, Pb, and Cd, respectively. The kinetics of PVA‐L‐CYS composite was accurately predicted by model of the pseudo‐first‐order kinetic. Thermodynamic prospects indicated a spontaneous, exothermic, and favorable uptake process at low temperatures. Efficient elution of the heavy metal ions from the overloaded composite was performed using 1 M HNO3. In accordance with WHO and FAO guidelines, the successful elimination of working metal ions from wastewater utilizing the PVA‐L‐CYS composite was demonstrated in a single cycle before discharge into the marine environment.

Effect of Multiantibiotic-Loaded Bone Cement on the Treatment of Periprosthetic Joint Infections of Hip and Knee Arthroplasties-A Single-Center Retrospective Study.

Two-stage septic revision is the prevailing method for addressing late periprosthetic infections. Using at least dual-antibiotic-impregnated bone cement leads to synergistic effects with a more efficient elution of individual antibiotics. Recent data on the success rates of multiantibiotic cement spacers in two-stage revisions are rare. We conducted a retrospective follow-up single-center study involving 250 patients with late periprosthetic hip infections and 95 patients with prosthetic knee infections who underwent septic two-stage prosthesis revision surgery between 2017 and 2021. In accordance with the antibiotic susceptibility profile of the microorganisms, a specific mixture of antibiotics within the cement spacer was used, complemented by systemic antibiotic treatment. All patients underwent preoperative assessments and subsequent evaluations at 3, 6, 9, 12, 18, and 24 months post operation and at the most recent follow-up. During the observation period, the survival rate after two-step septic revision was 90.7%. Although survival rates tended to be slightly lower for difficult-to-treat (DTT) microorganism, there was no difference between the pathogen groups (easy-to-treat (ETT) pathogens, methicillin-resistant staphylococci (MRS), and difficult-to-treat (DTT) pathogens). Furthermore, there were no differences between monomicrobial and polymicrobial infections. No difference in the survival rate was observed between patients with dual-antibiotic-loaded bone cement without an additional admixture (Copal® G+C and Copal® G+V) and patients with an additional admixture of antibiotics to proprietary cement. Employing multiple antibiotics within spacer cement, tailored to pathogen susceptibility, appears to provide reproducibly favorable success rates, even in instances of infections with DTT pathogens and polymicrobial infections.

3D-printed devices for continuous-flow lithium recovery of brines

Adsorption processes were widely used in the recovery of lithium from salt lakes. Fixed-bed continuous flow reactors were mainly used in adsorption due to their simple structure and stable reaction. 3D printing, due to its ability to greatly reduce the packing and fixed-bed reactor design cycle and cost, was gradually becoming as one of the key technologies for intelligent manufacturing of fixed beds. This work utilizes the advantages of digital preparation by 3D printing to address the low elution efficiency and adsorption capacity of titanium-based lithium-ion sieves (Ti-LIS) in fixed-bed lithium recovery. 3D printed fixed bed packing with high adsorption capacity were prepared using an in-situ growth strategy, acidic gel and reactor design were utilized to achieve efficient elution of Ti-LIS. The dynamic adsorption showed that the adsorption capacity of 3D printed fixed bed packing could reach 16 mg·g−1 and the elution efficiency can be increased from 78 % to 95 % with the same amount of acid.

Role of Calcium Chloride on the Eluting Process of Residual Ammonium from Weathered Crust Elution-Deposited Rare Earth Ore Tailings

A large amount of ammonium salt leaching agent will remain in the leaching site of weathered crust elution-deposited rare earth ore (WREOs). The release of residual ammonium (RA) will seriously affect the water system ecology of the mining area, and it is urgent to control it. In this paper, column eluting was used to simulate the eluting process of RA in rare earth (RE) ore tailings, and the effects of calcium chloride concentration, eluting temperature, liquid-solid ratio, eluent pH and eluent flow rate on the eluting process of RA in rare earth ore tailings were discussed. It was found that calcium chloride could effectively elute the RA from ore tailings. Eluting agent pH almost had no effect on the eluting process of RA in the pH range of 4–6, and a greater impact on it at pH 8. The flow rate could effectively enhance the elution efficiency. The optimum conditions were calcium ion concentration of 0.1 mol/L, liquid-solid ratio of 2:1, pH 4–6, flow rate of 0.6 mL/min and elution at room temperature. At this time, the elution efficiency of RA was 91.85%. The eluting process of RA in ore tailings was controlled by the inner particle diffusing according to the kinetic analysis. The reaction order was 0.368, and the activation energy of the reaction is 12.450 kJ/mol. This will provide a theoretical basis and technical support for the efficient eluting process of residual ammonium in the leaching site of WREOs.

In-Membrane Enrichment and Peptic Digestion to Facilitate Analysis of Monoclonal Antibody Glycosylation.

The number of therapeutic monoclonal antibodies (mAbs) is growing rapidly due to their widespread use for treating various diseases and health conditions. Assessing the glycosylation profile of mAbs during production is essential to ensuring their safety and efficacy. This research aims to rapidly isolate and digest mAbs for liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification of glycans and monitoring of glycosylation patterns, potentially during manufacturing. Immobilization of an Fc region-specific ligand, oFc20, in a porous membrane enables the enrichment of mAbs from cell culture supernatant and efficient elution with an acidic solution. Subsequent digestion of the mAb eluate occurred in a pepsin-modified membrane within 5 min. The procedure does not require alkylation and desalting, greatly shortening the sample preparation time. Subsequent LC-MS/MS analysis identified 11 major mAb N-glycan proteoforms and assessed the relative peak areas of the glycosylated peptides. This approach is suitable for the glycosylation profiling of various human IgG mAbs, including biosimilars and different IgG subclasses. The total time required for this workflow is less than 2 h, whereas the conventional enzymatic release and labeling of glycans can take much longer. Thus, the integrated membranes are suitable for facilitating the analysis of mAb glycosylation patterns.

Efficacy and safety of raltitrexed-eluting CalliSpheres® bead transarterial chemoembolization in patients with intermediate-stage hepatocellular carcinoma: a single-arm, prospective study.

The most common loadable chemotherapeutic drugs in drug-eluting bead transarterial chemoembolization (DEB-TACE) include doxorubicin, epirubicin, etc. CalliSpheres® beads have exhibited efficient loadability and eluting characteristics for raltitrexed as well as in vitro and animal experiments. However, the efficacy and safety of raltitrexed-loaded DEB-TACE in patients with intermediate-stage hepatocellular carcinoma (HCC) remain unclear. To assess the efficacy and safety of raltitrexed-loaded DEB-TACE in patients with intermediate-stage HCC. The study was conducted as a single-arm prospective study. This study was a prospective, single-arm trial conducted between June 2019 and June 2022. CalliSpheres® beads loaded with raltitrexed were used in the DEB-TACE procedure. The follow-up lasted for at least 1 year or until death. The primary endpoint was overall survival (OS), and the secondary endpoints were time to progression (TTP), progression-free survival (PFS), objective response rate (ORR), and adverse events (AEs). The 6-month ORR and disease control rates were 90.1% and 93.8%, respectively. The median OS was 33.0 months. The 1-, 2-, and 3-year survival rates were 95.1%, 82.1%, and 43.6%, respectively. Child-Pugh class and bilobar disease occurrence were identified as independent OS predictors. The median TTP and PFS were 22.7 and 19.8 months, respectively. Eleven (11.5%) patients experienced at least one grade 3 AE, and serious AEs were reported in five participants (5.2%). No patient experienced grade 4 or 5 AEs. Raltitrexed-loaded DEB-TACE is feasible, safe, and effective in patients with intermediate-stage HCC. This trial was registered at www.chictr.org.cn under the identifier: 1900024097 on 25 June 2019.

Therapeutic efficacy of vancomycin-loaded carbon fiber-reinforced polyetheretherketone hip stem for periprosthetic joint infection: A pilot study.

A carbon fiber-reinforced polyetheretherketone (CFR/PEEK) hip stem with a special antibiotic elution mechanism is under development to treat periprosthetic joint infection (PJI). The antibiotic elution characteristics of intramedullary implants were experimentally investigated, and the efficacy of revision surgery using a therapeutic stem in treating ovine PJI was examined. To evaluate elution characteristics, the intramedullary vancomycin-loaded CFR/PEEK cylindrical implants were inserted in the distal femur of nine sheep, and the vancomycin elution rate was measured at 2, 7, and 21 days. To evaluate therapeutic efficacy, the PJI model with staphylococcus aureus was attempted to create for five sheep. Moreover, the therapeutic vancomycin-loaded CFR/PEEK stem was implanted during one-stage revision surgery. Three weeks after revision surgery, the treatment efficacy was evaluated based on bacterial cultures and wound findings. In addition, the vancomycin elution rate from the stem was measured. On average, the cylindrical implants eluted approximately 70% vancomycin in 21 days. Of the five sheep attempting to create a PJI model, three were successfully infected with S. aureus as intended for verification of treatment efficacy. In all three joints, negative bacterial cultures and no purulence were observed 3 weeks after revision surgery. The vancomycin elution rates from the stems were >70%. Efficient elution of vancomycin was confirmed by the experimental implant inserted into the bone marrow and the stem in actual PJI treatment. Using a novel therapeutic stem with an antibiotic elution mechanism in one-stage revision surgery, successful treatment was demonstrated in all S. aureus-induced PJIs.

High Recovery Chromatographic Purification of mRNA at Room Temperature and Neutral pH.

Messenger RNA (mRNA) is becoming an increasingly important therapeutic modality due to its potential for fast development and platform production. New emerging RNA modalities, such as circular RNA, drive the need for the development of non-affinity purification approaches. Recently, the highly efficient chromatographic purification of mRNA was demonstrated with multimodal monolithic chromatography media (CIM® PrimaS), where efficient mRNA elution was achieved with an ascending pH gradient approach at pH 10.5. Here, we report that a newly developed chromatographic material enables the elution of mRNA at neutral pH and room temperature. This material demonstrates weak anion-exchanging properties and an isoelectric point of 5.3. It enables the baseline separation of mRNA (at least up to 10,000 nucleotides (nt) in size) from parental plasmid DNA (regardless of isoform composition) with both a NaCl gradient and ascending pH gradient approach, while mRNA elution is achieved in a pH range of 5-7. In addition, the basic structure of the novel material is a chromatographic monolith, enabling convection-assisted mass transfer of large RNA molecules to and from the active surface. This facilitates the elution of mRNA in 3-7 column volumes with more than 80% elution recovery and uncompromised integrity. This is demonstrated by the purification of a model mRNA (size 995 nt) from an in vitro transcription reaction mixture. The purified mRNA is stable for at least 34 days, stored in purified H2O at room temperature.

Developing the supraparticle technology for round window-mediated drug administration into the cochlea

The semi-permeable round window membrane (RWM) is the gateway to the cochlea. Although the RWM is considered a minimally invasive and clinically accepted route for localised drug delivery to the cochlea, overcoming this barrier is challenging, hindering development of effective therapies for hearing loss. Neurotrophin 3 (NT3) is an emerging treatment option for hearing loss, but its therapeutic effect relies on sustained delivery across the RWM into the cochlea. Silica supraparticles (SPs) are drug delivery carriers capable of providing long-term NT3 delivery, when injected directly into the guinea pig cochlea. However, for clinical translation, a RWM delivery approach is desirable. Here, we aimed to test approaches to improve the longevity and biodistribution of NT3 inside the cochlea after RWM implantation of SPs in guinea pigs and cats. Three approaches were tested (i) coating the SPs to slow drug release (ii) improving the retention of SPs on the RWM using a clinically approved gel formulation and (iii) permeabilising the RWM with hyaluronic acid. A radioactive tracer (iodine 125: 125I) tagged to NT3 (125I NT3) was loaded into the SPs to characterise drug pharmacokinetics in vitro and in vivo. The neurotrophin-loaded SPs were coated using a chitosan and alginate layer-by-layer coating strategy, named as ‘(Chi/Alg)SPs’, to promote long term drug release. The guinea pigs were implanted with 5× 125I NT3 loaded (Chi/Alg) SPs on the RWM, while cats were implanted with 30× (Chi/Alg) SPs. A cohort of animals were also implanted with SPs (controls). We found that the NT3 loaded (Chi/Alg)SPs exhibited a more linear release profile compared to NT3 loaded SPs alone. The 125I NT3 loaded (Chi/Alg)SPs in fibrin sealant had efficient drug loading (~5 μg of NT3 loaded per SP that weights ~50 μg) and elution capacities (~49% over one month) in vitro. Compared to the SPs in fibrin sealant, the (Chi/Alg)SPs in fibrin sealant had a significantly slower 125I NT3 drug release profile over the first 7 days in vitro (~12% for (Chi/Alg) SPs in fibrin sealant vs ~43% for SPs in fibrin sealant). One-month post-implantation of (Chi/Alg) SPs, gamma count measurements revealed an average of 0.3 μg NT3 remained in the guinea pig cochlea, while for the cat, 1.3 μg remained. Histological analysis of cochlear tissue revealed presence of a 125I NT3 signal localised in the basilar membrane of the lower basal turn in some cochleae after 4 weeks in guinea pigs and 8 weeks in cats. Comparatively, and in contrast to the in vitro release data, implantation of the SPs presented better NT3 retention and distribution inside the cochlea in both the guinea pigs and cats. No significant difference in drug entry was observed upon acute treatment of the RWM with hyaluronic acid. Collectively, our findings indicate that SPs and (Chi/Alg)SPs can facilitate drug transfer across the RWM, with detectable levels inside the cat cochlea even after 8 weeks with the intracochlear approach. This is the first study to examine neurotrophin pharmacokinetics in the cochlea for such an extended period of times in these two animal species. Whilst promising, we note that outcomes between animals were variable, and opposing results were found between in vitro and in vivo release studies. These findings have important clinical ramifications, emphasising the need to understand the physical properties and mechanics of this complex barrier in parallel with the development of therapies for hearing loss.

A simple and reliable isocratic high-performance chromatographic assay for the simultaneous determination of hydrophilic benzophenone-4 and lipophilic octocrylene in sunscreens.

This work aimed to develop a simple HPLC method for the simultaneous quantitative determination of the ultraviolet (UV) filters, hydrophilic benzophenone-4 and lipophilic octocrylene, in the presence of three other commonly used UV filters, avobenzone, octisalate and homosalate. Reverse-phased HPLC was performed on a C18 column. A scouting gradient was initially used to determine the approximate mobile phase composition required for efficient analyte elution and separation before further optimization. The assay was validated with regard to specificity, linearity, intra- and inter-day accuracy and precision, limits of detection and limits of quantification. An ultrasound dispersion extraction method for the UV filters from a commercial sunscreen was developed, and the extraction efficiencies from spiked samples were calculated. An acetonitrile-methanol-water mixture (20:67:13, v/v/v), where the water component contained 0.2% trifluoroacetic acid (v/v), was found to be the optimal mobile phase at a flow rate of 1.0 mL/min. The assay was linear between 1.0-100 μg/mL for both benzophenone-4 and octocrylene (both correlation coefficients were above 0.999). There was no interference from the excipients of the sunscreen nor from the three other UV filters. The intra- and inter-day accuracy was between 90.0-104.6% for both analytes. Extraction recoveries from a spiked commercial sunscreen were between 95.4 ± 2.1% to 98.5 ± 2.1% for benzophenone-4, and between 87.3 ± 2.3% and 98.9 ± 3.1% for octocrylene. All validation parameters were within the acceptance criteria set out in the International Council for Harmonization (ICH) guidelines. The HPLC assay showed the extracted quantities of benzophenone-4 and octocrylene from the commercial sunscreen closely matched claimed quantities. The developed isocratic HPLC method was suitable for simultaneously determining the hydrophilic benzophenone-4 and lipophilic octocrylene in the presence of other commonly used UV filters. Additionally, the extraction method was simple and effective for accurately quantifying the UV filters in a commercial sunscreen.

Discard the dross and select the essence: Purified orange peel with coupled alkali-acid hydrolysis as potential Cd(II)-capturing adsorbent

Turning agricultural wastes into green adsorbents is a compatible and cost-effective strategy for solid waste disposal and environmental pollution control. Herein, the discarded orange peel (OP) sample was purified via coupled alkali-acid hydrolysis and the obtained adsorbents were applied for eliminating toxic cadmium (Cd(II)) from the liquid phase. Based on the single factor experiments, the optimal Cd(II)-capturing adsorbent (i.e., OP-OH-H-25) was generated under the purification condition of soaking raw OP in 2.0 M NaOH solution at 25 °C followed by an acid hydrolysis treatment with 2.5 M HCl solution at 25 °C. Spectral analyses reflected the efficient elution of hemicellulose/pectin and lignin impurities, the transformation of cellulose type I to cellulose type II, and the exposure of sulfhydryl functional groups. The kinetics removal process of Cd(II) by OP-OH-H-25 reached equilibrium after 0.5 h with a removal efficiency of ∼88%. The maximum Cd(II) sorption capacity of OP-OH-H-25 (i.e., 124.90 mg/g) was higher than a series of previously reported adsorbents. The sorption behavior of Cd(II) onto the prepared OP-OH-H-25 material was mostly interfered with by the coexisting Mg2+, Ca2+, and heavy metals. The desorption data and spectroscopic analysis jointly pointed to the inner-sphere binding of Cd(II) with OP-OH-H-25 functional groups. Overall, the extracted OP-OH-H-25 sample could act as a promising decontaminant for the remediation of Cd(II)-polluted water environment.

Session IX:

Turning agricultural wastes into green adsorbents is a compatible and cost-effective strategy for solid waste disposal and environmental pollution control. Herein, the discarded orange peel (OP) sample was purified via coupled alkali-acid hydrolysis and the obtained adsorbents were applied for eliminating toxic cadmium (Cd(II)) from the liquid phase. Based on the single factor experiments, the optimal Cd(II)-capturing adsorbent (i.e., OP-OH-H-25) was generated under the purification condition of soaking raw OP in 2.0 M NaOH solution at 25 °C followed by an acid hydrolysis treatment with 2.5 M HCl solution at 25 °C. Spectral analyses reflected the efficient elution of hemicellulose/pectin and lignin impurities, the transformation of cellulose type I to cellulose type II, and the exposure of sulfhydryl functional groups. The kinetics removal process of Cd(II) by OP-OH-H-25 reached equilibrium after 0.5 h with a removal efficiency of ∼88%. The maximum Cd(II) sorption capacity of OP-OH-H-25 (i.e., 124.90 mg/g) was higher than a series of previously reported adsorbents. The sorption behavior of Cd(II) onto the prepared OP-OH-H-25 material was mostly interfered with by the coexisting Mg2+, Ca2+, and heavy metals. The desorption data and spectroscopic analysis jointly pointed to the inner-sphere binding of Cd(II) with OP-OH-H-25 functional groups. Overall, the extracted OP-OH-H-25 sample could act as a promising decontaminant for the remediation of Cd(II)-polluted water environment.

Sorption and desorption performance of La3+/Bi3+ by surface-modified activated carbon for potential application in medical 225Ac/213Bi generators

213Bi is a promising candidate for targeted alpha therapy.However, the search for suitable materials to separate clinically relevant activities of 213Bi from its parent 225Ac remains an ongoing yet critical challenge. This study evaluates sulfonated Norit CA1 (activated carbon) materials (SNCAs) as alternative sorbents for use in 225Ac/213Bi radionuclide generators and gives an overview of their simultaneous sorption/desorption performance towards La3+ and Bi3+. The SNCAs exhibit high and selective uptake of Bi3+ in the presence of NaNO3 at low pH values. In contrast, the sorption performance for La3+ (which was certified as a non-radioactive surrogate for 225Ac3+) onto SNCAs was more sensitive to the salt concentrations and pH values. The desorption results show that an efficient elution of Bi3+ from SNCAs is achieved using HCl, NaI or NaCl solutions. The experimental sorption data are analysed by various kinetic and isotherm models. The SNCAs exhibit high radiolytic stability, fast sorption kinetics, good durability and recyclability for La3+/Bi3+. This research indicates that SNCAs with sufficient oxygen-containing groups are promising sorbents for the separation of 213Bi in an inverse 225Ac/213Bi radionuclide generator.

In vitro elution characteristics of antibiotic-loaded polymethylmethacrylate cement and a calcium sulfate bone substitute using staphylococci isolated from orthopedic implant-associated infections.

Polymethylmethacrylate (PMMA) remains the gold standard antibiotic carrier in the management of osteomyelitis. However, biodegradable ceramic carriers may exhibit more efficient antibiotic elution properties. Through zone of inhibition (ZOI) testing and biofilm killing assays, we assessed the in vitro elution efficacy of vancomycin released from calcium sulfate (PG-CSH) and PMMA beads as carriers on clinical strains of Staphylococcus aureus and Staphylococcus epidermidis, which were isolated from sonication fluid of orthopedic implant-associated infections. Overall, vancomycin-loaded PMMA and PG-CSH beads showed potency (ZOI above 4cm2 ) for up to 14 days against ATCC and clinical strains. Vancomycin-loaded PG-CSH beads displayed higher rates, exhibited a more stable antibiotic elution, had greater impacts on bacterial colony-forming unit counts and produced higher ZOIs; additionally, statistically significant differences (Student's t test) were observed in different time sets during the experiment. In the biofilm killing assay, PG-CSH loaded with vancomycin resulted in more bacterial deaths. In conclusion, in the present study, both PG-CSH and PMMA beads acted as good carriers, but greater antimicrobial elution and biofilm bacterial killing were observed with PG-CSH than PMMA. Future in vitro research should focus on testing other difficult-to-treat clinical strains, including multidrug resistant coagulase-negative staphylococci and Gram-negative bacilli.

Localized Instillation Enables In Vivo Screening of Targeting Peptides Using One-Bead One-Compound Technology.

The One-Bead One-Compound (OBOC) library screening is an efficient technique for identifying targeting peptides. However, due to the relatively large bead size, it is challenging for the OBOC method to be applied for in vivo screening. Herein, we report an in vivo Localized Instillation Beads library (LIB) screening method to discover targeting peptides with the OBOC technique. Inspired by localized instillation, we constructed a cavity inside of a transplanted tumor of a mouse. Then, the OBOC heptapeptide library was injected and incubated inside the tumor cavity. After an efficient elution process, the retained beads were gathered, from which three MDA-MB-231 tumor-targeting heptapeptides were discovered. It was verified that the best peptide had 1.9-fold higher tumor accumulation than the commonly used targeting peptide RGD in vivo. Finally, two targeting proteins were discovered as potential targets of our targeting peptide to the MDA-MB-231 tumor. The in vivo LIB screening method expands the scope of OBOC peptide screening applications to discover targeting peptides in vivo feasibly and reliably.

Spy&IAC enables specific capture of SpyTagged proteins for rapid assembly of plug-and-display nanoparticle vaccines

From modular vaccine production to protein assembly on nanoparticles, the SpyCatcher/SpyTag system provides a convenient plug-and-display procedure. Here, we established a general-purpose immunoaffinity chromatography (IAC) method for SpyTagged proteins (Spy&IAC). SpyTags are displayed on the surface of nanoparticles to induce high-affinity monoclonal antibodies, allowing the specific capture of the target protein. Taking the key core antigenic regions of two coronaviruses that are currently more threatened in the field of human and animal diseases, the nucleocapsid (N) protein of SARS-CoV-2 and the COE protein of porcine epidemic diarrhea virus (PEDV) as model proteins, a purification model with SpyTag at the N-terminal or C-terminal expressed in E. coli or mammalian cells was constructed. After the efficient elution of Spy&IAC, the final yield of several proteins is about 3.5–15 mg/L culture, and the protein purity is above 90 %. Purification also preserves the assembly function and immunogenicity of the protein to support subsequent modular assembly and immunization programs. This strategy provides a general tool for the efficient purification of SpyTagged proteins from different expression sources and different tag positions, enabling the production of modular vaccines at lower cost and in a shorter time, which will prepare the public health field for potential pandemic threats.

Immunostimulatory effects of a subcritical water extract of Ganoderma.

Ganoderma, a medicinal mushroom with various physiological activities, has been extensively investigated regarding its effectiveness. The aim of the present study was to examine the effects of a subcritical water extract of Ganoderma (SWEG) on the immune system. The use of subcritical water with a higher temperature and pressure than hot water allows efficient elution of components from natural products. As an evaluation of the effectiveness of SWEG, a cell proliferation and a cell differentiation test were carried out using A-6 cells, a model of hematopoietic stem cells. Furthermore, an oral administration test in mice was conducted to examine the effects of SWEG on the number and function of immune cells. As a result, SWEG was revealed to promote both self-renewal and differentiation into immune cells such as T cells and natural killer (NK) cells in experiments with A-6 cells. These results were not obtained in experiments using hot water extract of Ganoderma lucidum and Ganoderma sinense. The oral administration test in mice demonstrated that SWEG increased hematopoietic precursor cells, immature B cells, and NK cells in the bone marrow, and T cells in the thymus. In addition, SWEG enhanced the immune functions in the spleen by promoting granzyme B expression and NK cell activity. SWEG was demonstrated to be a food material that acts on HSCs and regulates immunity in vivo.

Quantification of Pitolisant: A RP - HPLC study

Objective An attempt was made to develop a simple, isocratic and robust reverse phase high performance liquid chromatographic method for Pitolisant. Material and methods An Inertsil octa decyl silane column and photo diode array detector were employed for the quantification of Pitolisant at 268 nm using a mobile phase consisting of acetonitrile and 0.1 % v/v formic acid (90:10 % v/v) at a flow rate of 1.0 mL/min. for the efficient elution of Pitolisant. Results The developed method was validated for various parameters and the linearity range was observed at 25-150 µg/mL. The correlation coefficient, limit of detection and limit of quantification calculated from the calibration curve were 0.9998, 0.3 µg/mL and 1.0 µg/mL respectively. The mean percentage recovery was 99.9 %. The formulation was subjected to stress at various conditions resulting in minor degradants at different retention times. Conclusion An isocratic, fast, robust reverse phase – high performance liquid chromatographic method was developed which can be exploited for the estimation of Pitolisant in bulk and tablets. Key words: Pitolisant, High performance liquid chromatography, Photo diode array detector, Limit of detection, Degradation studies