Large-scale Purification Research Articles

Photothermal Synergistic Hydrogen Production via a Fly-Ash-made Interfacial Vaporific System.

Employing UV-vis spectrum for hydrogen generation and vis-IR spectrum to elevate reaction temperatures and induce phase transitions effectively enhances yield and purifies water, demonstrating a judicious strategy for solar energy utilization. This study presents an interfacial photothermal water splitting system that utilizes all-inorganic, economical industrial by-products known as fly ash cenospheres (FAC) for solar-driven hydrogen generation. In this system, the yield reaches 254.8µmol h-1cm-1, representing an 89% augmentation compared to that of the three-phase system. In situ experiments, combined with theoretical calculation, reveal the system's robust light absorption capacity, facilitating rapid gas separation, thus improves the solar-to-hydrogen (STH) efficiency. Furthermore, the system demonstrates strong performance in turbid water and scalability for expansive applications, achieving a hydrogen yield exceeding 50 L h-1 m-2 from various water sources. Facilitating large-scale hydrogen production and water purification, it thereby establishing its potential as a viable solution for sustainable energy generation.

Macroporous resins combined with UPLC-QTRAP-MS/MS for the purification and identification of 3-deoxyanthocyanidins from Sorghum bicolor L. Moench and their bioactivities.

3-Deoxyanthocyanidins (3-DAS) are the unique flavonoid compounds in sorghum. The majority of research to date have concentrated on the biological activity and extraction of 3-DAS from sorghum and lacked systematic purification and identification investigations. Herein, a facile method for the purification of sorghum 3-DAS from the acidic methanol solution crude extract using macroporous resins (MARs) was proposed and investigated in this work. Based on the experimental data, commercial MAR AB-8 was selected to achieve the purification of sorghum 3-DAS. The adsorption-desorption experiment confirmed the optimal procedure. With this procedure, the total purity of 3-DAS was 19.32%, and 3-DAS content in the purified product was 8.4 times higher than that in the crude extract. UPLC-QTRAP-MS/MS identification indicated that the purified 3-DAS product was mainly composed of apigeninidin, luteolinidin, and their methoxy derivatives. Besides, the purified product exhibited better bioactivities than the crude extract, including the scavenging activities for DPPH, ABTS+, and hydroxyl radicals, the total antioxidant capacity, and the inhibitory effects for α-amylase, α-glucosidase, and lipase. This work provides a potential way for the large-scale purification of bioactive sorghum 3-DAS from crude extract, which is significant to deepen the applications of sorghum 3-DAS in the food nutrition and pharmaceutical industries.

Purification of plasmid DNA using a novel two stage chromatography process

The chromatography process of large-scale plasmid purification with high efficiency and low cost has always been a major challenge. We established a two-step plasmid chromatography purification process combining multimodal and thiophilic chromatography with an overall chromatography yield of nearly 70%. Capto Core 700, a multimodal core–shell particle, was firstly used to remove the impurities from the crude lysate. The effects of different experimental conditions on chromatography recovery and impurity removal were screened. Compared to conventional size exclusion chromatography, the sample load and flow rate of this step were enhanced by 40-fold and 5-fold, respectively, while maintaining a 90% yield. For the thiophilic chromatography (Capto PlasmidSelect), the method of Design of Experiments (DoEs) was used to study the influence of parameters on the results. The effects of ammonium sulfate concentration, sodium chloride concentration and flowrate in the elution phase were studied and optimized with a central composite design model consisting of 17 experiments. The versatility of this process was demonstrated by successfully purifying three different lentiviral packaging plasmids (pLP1, pLP2 and pLP/VSVG) and the target plasmid containing green fluorescent protein (GFP). Purified plasmids consistently achieved a supercoiled purity of at least 90% with endotoxin levels below 5 EU/mg. Lentiviral vectors packaged using these plasmids exhibited high infectious titers of 1 × 107 TU/mL, thereby verifying the process applicability for diverse plasmid purification requirements.

Scale-up construction of stable multifunctional hydrogel interfaces for large-scale purification of complex oil-water emulsions and oil recovery

Inspired by the multi-level structure of grass clumps in nature, a novel filter with plexiform-structured hydrogel interface was constructed using sepiolite-derived silica nanofiber (SiNF) as the supporter and crosslinked polyvinyl alcohol (cl-PVA) hydrogel as the coating. Experimental test, DFT and MD calculations have confirmed that the addition of SiNF can not only enhance oil-water separation efficiency, but also improve the stability of hydrogel coating. The hydrogel interface with excellent stability and superhydrophilic/underwater superoleophobicity can be manufactured on a large copper mesh (1 m × 1.2 m) to achieve large-scale production. The surface-engineered mesh (named cl-PVA/SiNF@Ag-Cu) can be assembled on a self-designed equipment for continuous purification of emulsion wastewater (processing capacity: 576.00 L/day), achieving a high separation efficiency of 99.7 % for complex oily emulsion only under the action of gravity, and can simultaneously recover oils. After being treated under extreme conditions such as strong acid/alkali, high/low temperature (100 °C, 200 °C, and −18 °C), high salt concentration, sandpaper wear, and long-term aging, the surface structure of cl-PVA/SiNF@Ag-Cu filter remains stable. The antifouling, antibacterial, and anticorrosion capabilities of the filter give it the potential for long-term and large-scale purification processes. Planting and breeding experiments have confirmed that purified water is harmless to animals and plants.

Biophysical characterization and structural insights of leptospiral complement regulator-acquiring protein A

Many pathogens establish a successful infection by evading the host complement system, an essential arm of innate immunity. Pathogenic Leptospira is reported to escape complement-mediated killing by recruiting the host complement regulators by lipoproteins or outer surface proteins. One of the outer surface proteins, Leptospiral complement regulator-acquiring protein A (LcpA), is known to recruit complement regulators, C4b-binding protein (C4BP), and Factor H (FH) on the bacterial surface. Mapping of interacting domains from C4BP and FH with the LcpA has already been reported. However, the region or structural part of the LcpA mediating the interaction is not known yet. Here, we report cloning, expression, refolding and purification of recombinant LcpA from an inclusion body of E. coli heterologous expression system. We also demonstrate the biophysical characterization of recombinant LcpA and reveal its secondary structure contents. Moreover, the protein displays a moderate thermostability. The change of intrinsic fluorescence and CD spectra demonstrate a change in the secondary structure of protein due to binding with Zn2+ ions. Molecular docking of LcpA with the complement regulators displays important interface residues from both the individual counterparts. Molecular dynamic simulation analysis demonstrates the stability of interactions between LcpA and C4BP. In our understanding, this is the first report on the large-scale purification of LcpA through refolding experiments and biophysical characterization of LcpA. This study may provide additional information on the structural basis of binding with the complement regulators.

Bioengineered lubricin alters the lubrication modes of cartilage in a dose-dependent manner.

The low friction nature of articular cartilage has been attributed to the synergistic interaction between lubricin and hyaluronic acid in the synovial fluid (SF). Lubricin is a mucinous glycoprotein that lowers the boundary mode coefficient of friction of articular cartilage in a dose-dependent manner. While there have been multiple attempts to produce recombinant lubricin and lubricin mimetic cartilage lubricants over the last two decades, these materials have not found clinical use due to challenges associated with large scale production, manufacturing, and purification. Recently, a novel method using codon scrambling was developed to produce a stable, full-length bioengineered equine lubricin (eLub) in large reproducible quantities. While preliminary frictional analysis of eLub and other recombinantly produced forms revealed they can lubricate cartilage, a complete tribological characterization is lacking, with previous studies evaluating the friction coefficient only at a single dose or a single speed. The objective of this study was to analyze the dose-dependent tribological properties of eLub using the Stribeck framework of tribological analysis. Recombinantly produced eLub at doses greater than 1.5 mg/mL exhibits friction coefficients on par with healthy bovine SF, and a maximal 5 mg/mL dose exhibits a nearly 50% lower friction coefficient than healthy SF. eLub also modulates the shift in lubrication mode of the cartilage from the high friction boundary mode to the low friction minimum mode at high concentrations.

For catching-by-polymerization oligo purification: scalable synthesis of the precursors to the polymerizable tagging phosphoramidites

The catching-by-polymerization (CBP) oligodeoxynucleotide (oligo or ODN) purification method has been demonstrated suitable for large-scale, parallel, and long oligo purification. The authenticity of the oligos has been verified via DNA sequencing, and gene construction and expression. A remaining obstacle to the practical utility of the CBP method is affordable polymerizable tagging phosphoramidites (PTPs) that are needed for the method. In this article, we report scalable synthesis of the four nucleoside (dA, dC, dG and T) precursors to the PTPs using a route having five steps from inexpensive starting materials. The overall yields ranged from 21% to 35%. The scales of the synthesis presented here are up to 2.1 grams of the precursors. Because the syntheses are chromatography-free, further scaling up the syntheses of the precursors have become more feasible. With the precursors, the PTPs can be synthesized in one step using standard methods involving a chromatography purification.

Development of cost-effective cellulose-based bilayer hybrid composite membranes for CO2 separation in biogas purification

CO2 is the main pollutant in biogas, reducing its calorific value. Among various technological methods to eliminate carbon dioxide from biogas, membrane separation technology stands out for large-scale industrial biogas purification due to its advantages. The selection of membrane material and preparation process are key factors in membrane separation technology. In this study, a premixing process was initially used to blend different masses of packed molecular sieves TS-1 (or ZSM-5) and cellulose derivatives (ethyl cellulose, cellulose acetate) separately. These mixtures were then coated onto porous PVDF substrates using a coating process to create various bilayer hybrid composite membranes. Among these, PVDF/EC-ZSM-5 (containing 15 % ZSM-5) bilayer hybrid composite membrane is the most fitting. For CO2/CH4 gas mixtures, the gas selectivity of this membrane surpassed Robeson's 1991 standard line (the CO2 permeability was 597.48 Barrer, and the CO2/CH4 selectivity was 9.14). Overall, this composite membrane, made for the first time from PVDF, ZSM-5, and EC, is expected to be a promising CO2 selective separation membrane material for biogas purification in large-scale industrial processes due to its simple production process.

Innovative Charge-Tuning for Highly Dispersed Pt Catalysts: Achieving Deep CO Removal in Industrial H2 Purification for Fuel Cells.

Proton exchange membrane fuel cells have strict requirements for the CO concentration in H2-rich fuel gas. Here, from the perspective of industrial practicability, a highly dispersed Pt catalyst (2-4 nm) supported on activated carbon (AC), which was modified by electronic promoters (K+) and structural promoters (isopropanol), is studied in detail. Compared with traditional metal oxide supports, the K-Pt/AC catalysts, which benefit from the tuned charge distribution, achieve a significant reduction of CO (from 1% to <0.1 ppb) under H2-rich conditions and show potential for used in large-scale industrial hydrogen purification. Experimental results and theoretical calculations reveal that the K atom, with its lower electronegativity, contributes to the shift of surface Pt2+ to a lower binding energy due to the presence of oxygen species on the AC surface. This facilitates oxygen activation and accelerates desorption of the CO2 product, thereby accelerating the reaction process and enabling the deep removal of CO in a hydrogen-rich atmosphere.

Effective photocatalytic degradation of sulfamethoxazole using PAN/SrTiO3 nanofibers

Freshwater scarcity driven by pollution and climate change necessitates the development of effective water purification methods. There is an urgent need for innovative solutions that can efficiently remove contaminants from water sources. A effective material for the photocatalytic degradation of organic sulfonamide antibiotics from water was developed in this study. A composite of electrospun strontium titanate (SrTiO3) and carbon fibers polyacrylonitrile/strontium titanate (PAN/SrTiO3) was employed for the photocatalytic degradation of sulfamethoxazole (SMX). Scanning Electron Microscopy/Energy Dispersive Spectroscopy (SEM-EDS), Transmission electron microscopy (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS) were used to characterise the composite. The fibers had a thickness of ~250 nm and uniformly dispersed PAN and SrTiO3 particles. A PAN/SrTiO3 composite with a 1:1 mass ratio (20 mg) demonstrated excellent performance, achieving over 90 % degradation at pH 7 and a contaminant concentration of 5 mg/L under ultraviolet (UV) light irradiation during 60 min. The composite was tested for the photodegradation of the major organic pollutant (SMX) in distilled and river water. In river water, the degradation efficiency exceeded 80 % at pH 7 because of the formation of secondary radicals involved in antibiotic degradation. SMX degradation was described using a pseudo-first order kinetic equation using the Langmuir-Hinshelwood model. These results underscore the novelty and significance of the PAN/SrTiO3 composite and demonstrate its high efficiency and potential for large-scale water purification applications.

Force-enhanced sensitive and specific detection of DNA-intercalative agents directly from microorganisms at single-molecule level.

Microorganisms can produce a vast array of bioactive secondary metabolites, including DNA-intercalating agents like actinomycin D, doxorubicin, which hold great potential for cancer chemotherapy. However, discovering novel DNA-intercalating compounds remains challenging due to the limited sensitivity and specificity of conventional activity assays, which require large-scale fermentation and purification. Here, we introduced the single-molecule stretching assay (SMSA) directly to microbial cultures or extracts for discovering DNA-intercalating agents, even in trace amounts of microbial cultures (5 μl). We showed that the unique changes of dsDNA incontour length and overstretching transition enable thespecific detection of intercalators from complex samples without the need forextensive purification. Applying force to dsDNA also enhanced the sensitivity by increasing both the binding affinity Ka and the quantity of ligands intercalation, thus allowing thedetection of weak intercalators, which are often overlooked using traditional methods. We demonstrated the effectiveness of SMSA,identified two DNA intercalator-producing strains: Streptomyces tanashiensis and Talaromyces funiculosus, and isolated three DNA intercalators: medermycin, kalafungin and ligustrone B. Interestingly, both medermycin and kalafungin, classified as weak DNA intercalators (Ka ∼103 M-1), exhibited potent anti-cancer activity against HCT-116 cancer cells, with IC50 values of 52±6and 70±7 nM, respectively.

Bisphenol A in Water Systems: Risks to Polycystic Ovary Syndrome and Biochar-Based Adsorption Remediation: A Review.

Access to clean and safe water sadly remains an issue in the 21st century. Water reservoirs, whether groundwater or surface water, are routinely contaminated by various harmful Emerging Contaminants (ECs). One of most prevalent pollutants among these pollutants is Bisphenol A, which is classified as an Endocrine Disrupting Compound (EDC). This substance adversely interferes with the endocrine system, primarily by mimicking estrogen, and has been considered a potential contributor to Polycystic Ovary Syndrome (PCOS) with 82.70% of 1,391 women studied showing a positive correlation between BPA exposure and PCOS. PCOS is currently the most prevalent endocrine disorder affecting women of reproductive age; however, its pathogenesis remains unclear, complicating diagnosis and subsequently patient care. In this review, these topics are thoroughly examined, with particular emphasis on biochar, a new promising method for large-scale water purification. Biochar, derived from various organic waste materials, has emerged as a cost-effective substance with remarkable adsorption properties achieving up to 88% efficiency over four cycles of reuse, similar to that of activated carbon. This review interrogates the suitability of biochar for counteracting the issue of EDC pollutants.

Large-scale purification of a deprotected macrocyclic peptide by supercritical fluid chromatography (SFC) integrated with liquid chromatography in discovery chemistry

A macrocyclic peptide A was successfully purified in large quantities (∼30 g) in >95 % purity by an integrated two-step orthogonal purification process combining supercritical fluid chromatography (SFC) with medium-pressure reverse-phase liquid chromatography (MP-RPLC). MP-RPLC was used to fractionate the crude peptide A, remove unwanted trifluoroacetic acid (TFA) originating from the peptide A cleavage off the resin, and convert the peptide A into ammonium acetate salt form, prior to the final purification by SFC. A co-solvent of methanol/acetonitrile containing ammonium acetate and water in CO2 was developed on a Waters BEH 2-Ethylpyridine column. The developed SFC method was readily scaled up onto a 5 cm diameter column to process multi-gram quantities of the MP-RPLC fraction to reach > 95 % purity with a throughput/productivity of 0.96 g/h. The incorporation of SFC with MP-RPLC has been demonstrated to have a broader application in other large-scale polypeptide purifications.

Breakthrough in High-Efficiency Photocatalytic Degradation of Acebutolol by Advanced Binary CeO2-MnO2 Oxide System.

The sustainable catalytic efficacy of transition metal oxides (TMO) and rare earth element-based oxides positions them as pivotal materials for effectively treating contaminated wastewater. This study successfully synthesized a series of Ce@MnO2 photocatalysts using a straightforward hydrothermal method. These photocatalysts were thoroughly characterized for their optical properties, structural morphology, and phase purity. Among the synthesized materials, the Ce@MnO2 (40:60) exhibited the highest photocatalytic activity for the degradation of Acebutolol (ACB), achieving a remarkable degradation efficiency of 92.71% within 90 min under visible light irradiation. This superior performance is attributed to the increased presence of active species and the efficient separation of photogenerated carriers. Additionally, the photocatalytic reaction mechanism was elucidated, highlighting the catalyst's surface charge properties which significantly enhanced performance in a solution with pH 8. The outstanding photo-response in the visible spectrum renders this method not only cost-effective but also environmentally benign, presenting a promising approach for large-scale water purification.

Soluble and insoluble expression of recombinant human interleukin-2 protein using pET expression vector in Escherichia coli

Interleukin-2 has emerged as a potent protein-based drug to treat various cancers, AIDS, and autoimmune diseases. Despite its immense requirement, the production procedures are inefficient to meet the demand. Therefore, efficient production procedures must be adopted to improve protein yield and decrease procedural loss. This study analyzed cytoplasmic and periplasmic IL-2 expression for increased protein yield and significant biological activity. The study is focused on cloning IL-2 into a pET-SUMO and pET-28a vector that expresses IL-2 in soluble form and inclusion bodies, respectively. Both constructs were expressed into different E. coli expression strains, but the periplasmic and cytoplasmic expression of IL-2 was highest in overnight culture in Rosetta 2 (DE3). Therefore, E. coli Rosetta 2 (DE3) was selected for large-scale production and purification. Purified IL-2 was characterized by SDS-PAGE and western blotting, while its biological activity was determined using MTT bioassay. The results depict that the periplasmic and cytoplasmic IL-2 achieved adequate purification, yielding 0.86 and 0.51 mg/mL, respectively, with significant cytotoxic activity of periplasmic and cytoplasmic IL-2. Periplasmic IL-2 has shown better yield and significant biological activity in vitro which describes its attainment of native protein structure and function.

A self-floating integrated hydrogel evaporator with efficient salt resistance and thermal localization for efficient solar water desalination

Solar-driven interfacial evaporation technology was considered a promising way to effectively alleviate the global freshwater crisis. However, it is still a major challenge to design evaporators with efficient thermal management, salt resistant and self-floating ability to realize efficient clean water production. Here, we design a double-layer hydrogel evaporator with self-floating system. The evaporator is prepared from expanded polystyrene board, poly (vinyl alcohol), polyvinylpyrrolidone, agar and conductive carbon black. The structural design of the evaporator improves water transport capacity of the evaporation system and effectively reduces heat loss, so that the evaporator has efficient salt resistance and thermal localization. The evaporator demonstrates an outstanding evaporation rate and evaporation efficiency of 2.77 kg m-2h−1 and 94.25 % under 1 kW m−2 illumination. Additionally, the evaporator exhibits good stability in the 10-cycle solar evaporation experiment. Furthermore, the evaporator exhibits excellent salt resistance in 3.5 wt% NaCl solution during continuous evaporation for 9 h at 1 kW m−2 illumination. Moreover, the purification rates of the organic wastewater 4-nitrophenol and methylene blue by the evaporator reached 99.8 % and 100 %, respectively. This evaporator has stable and efficient capability of solar water purification, providing a new way for large-scale solar desalination and water purification.

Polymeric monolithic columns based on natural wood for rapid purification of targeted protein

With multiscale hierarchical structure, wood is suitable for a range of high-value applications, especially as a chromatographic matrix. Here, we have aimed to provide a weak anion-exchange polymeric monolithic column based on natural wood with high permeability and stability for effectively separating the targeted protein. The wood-polymeric monolithic column was synthesized by in situ polymerization of glycidyl methacrylate and ethylene glycol dimethacrylate in wood, and coupled with diethylaminoethyl hydrochloride. The wood-polymeric monolithic column can be integrated with fast-protein liquid chromatography for large-scale protein purification. According to the results, the wood-polymeric monolithic column showed high hydrophilicity, permeability and stability. Separation experiments verified that the wood-polymeric monolithic column could purify the targeted protein (spike protein of SARS-COV-2 and ovalbumin) from the mixed proteins by ion exchange, and the static adsorption capacity was 33.04 mg mL−1 and the dynamic adsorption capacity was 24.51 mg mL−1. In addition, the wood-polymerized monolithic column had good stability, and a negligible decrease in the dynamic adsorption capacity after 20 cycles. This wood-polymerized monolithic column can provide a novel, efficient, and green matrix for monolithic chromatographic columns.

Large-scale separation and purification of exosomes using ionexchange chromatography

Large-scale separation and purification of exosomes using ionexchange chromatography

Floating MIL-88A(Fe)@expanded perlites catalyst for continuous photo-Fenton degradation toward tetracyclines under artificial light and real solar light

In this work, MIL-88A(Fe) was immobilized onto the expanded perlites to fabricate the floating MIL-88A(Fe)@expanded perlites (M@EP) catalyst via high throughput batch synthesis method under room temperature. The as-prepared M@EP could efficiently activate H2O2 to achieve 100% tetracycline antibiotics (TCs) removal under both artificial low power UV light (UVL) and real sunlight (SL) irradiation. The toxicological evaluation, growth experiment of mung beans and antimicrobial estimation revealed the decreasing aquatic toxicity of the TCs intermediates compared to those of the pristine TCs. A self-designed continuous bed reactor was employed to investigate the long-term operation of the M@EP. The findings demonstrated that the antibiotics mixture can be continuously degraded up to 7 days under UVL and 5 daytimes under SL irradiation, respectively. More importantly, ca. 76.9% and 81.6% of total organic carbon (TOC) removal efficiencies were accomplished in continuous bed reactor under UVL and SL irradiation, respectively. This work advances the immobilized MOFs on floating supports for their practical application in large-scale wastewater purification through advanced oxidation processes. Environmental implicationThis work presented the high throughput production and photo-Fenton degradation application of floating MIL-88A(Fe)@expanded perlites (M@EP). Three tetracycline antibiotics (TCs) were selected as model pollutants to test the degradation ability of M@EP in batch experiment and continuous operation under artificial light and solar light. The complete TCs degradation could be accomplished in self-designed device up to 7 d under UV light and 5 d under real solar light. This work tapped a new door to push MOFs-based functional materials in the real water purification.

Plasmodium RNA triphosphatase validation as antimalarial target

Target-based approaches have traditionally been used in the search for new anti-infective molecules. Target selection process, a critical step in Drug Discovery, identifies targets that are essential to establish or maintain the infection, tractable to be susceptible for inhibition, selective towards their human ortholog and amenable for large scale purification and high throughput screening. The work presented herein validates the Plasmodium falciparum mRNA 5’ triphosphatase (PfPRT1), the first enzymatic step to cap parasite nuclear mRNAs, as a candidate target for the development of new antimalarial compounds. mRNA capping is essential to maintain the integrity and stability of the messengers, allowing their translation. PfPRT1 has been identified as a member of the tunnel, metal dependent mRNA 5′ triphosphatase family which differs structurally and mechanistically from human metal independent mRNA 5′ triphosphatase. In the present study the essentiality of PfPRT1 was confirmed and molecular biology tools and methods for target purification, enzymatic assessment and target engagement were developed, with the goal of running a future high throughput screening to discover PfPRT1 inhibitors.