Purification Platform Research Articles

Clear insight into complex multimodal resins and impurities to overcome recombinant protein purification challenges: A review.

Increasing attention has been paid to the purity of therapeutic proteins imposing extensive costs and challenges to the downstream processing of biopharmaceuticals. One of the efforts, that has been exerted to overcome such limitations, was developing multimodal or mixed-mode chromatography (MMC) resins for launching selective, orthogonal, non-affinity purification platforms. Despite relatively extensive usage of MMC resins, their real potential and fulfillment have not been extensively reviewed yet. In this work, the explanation of practical and key aspects of downstream processing of recombinant proteins with or without MMC resins was debated, as being useful for further purification process development. This review has been written as a step-by-step guide to deconvolute both inherent protein purification and MMC complexities. Here, after complete elucidation of the potential of MMC resins, the effects of frequently used additives (mobile phase modifiers) and their possible interactions during the purification process, the critical characteristics of common product-related impurities (e.g., aggregates, charge variants, fragments), host-related impurities (e.g., host cell protein and DNA) and process related impurities (e.g., endotoxin, and viruses) with solved or unsolved challenges of traditional and MMC resins have been discussed. Such collective experiences which are reported in this study could be considered as an applied guide for developing successful downstream processing in challenging conditions by providing a clear insight into complex MMC resins and impurities.

Quality by design for mRNA platform purification based on continuous oligo-dT chromatography

Oligo-deoxythymidine (Oligo-dT) ligand-based affinity chromatography is a robust method for purifying mRNA drug substances within the manufacturing process of mRNA-based products, including vaccines and therapeutics. However, the conventional batch mode of operation for oligo-dT chromatography has certain drawbacks that reduce this process’s productivity. Here, we report a new continuous oligo-dT chromatography process for the purification of in vitro transcribed mRNA, which reduces losses, improves the efficiency of oligo-dT resin use, and intensifies the chromatography process. Furthermore, the quality by design (QbD) framework was used to establish a design space for the newly developed method. The optimisation of process parameters, including salt type, salt concentration, load flow rate and mRNA load concentration both in batch and the continuous mode, achieved >90% yield (mRNA recovery) along with >95% mRNA integrity and >99% purity. The productivity of continuous chromatography was estimated to be 5.75-fold higher, and the operating cost was estimated 15% lower, when compared to batch chromatography. Furthermore, the QbD framework was further used to map the relationship between critical quality attributes (CQAs) and key performance indicators (KPIs) as a function of critical process parameters (CPPs) and critical material attributes (CMAs).

A SARS-CoV-2 peptide antigen purified from bacteria and displayed in a high-density repetitive manner on a virus-like particle could generate anti-SARS-CoV-2 neutralizing antibodies unlike free peptide

SARS-CoV-2 is an enveloped virus. Proteins of the lipid based envelope are not rigidly arranged as compared to non-enveloped viruses lacking lipid based outer layer. Rigidly arranged multivalent display has been reported to be important for potent immune stimulation. We assembled himeric virus-like-particle (VLP) where the core of the particle is composed of the coat protein of Acinetobacter phage. Peptide-based antigen from receptor binding domain (RBD) of SARS-CoV-2 spike protein has been displayed on the coat based VLP matrix using spy-tag/spy-catcher split inteine conjugation system that allows spontaneous, irreversible isopeptide bond formation. Both the matrix as well as peptide have been purified from bacteria which is an easy platform for protein purification. We used the closed state of spike trimer for epitope mapping as this is the conformation of the spike that the immune system visualizes unlike the open state that appears when the virus tries to attach to receptor. Mice based immunization studies were used to test immunogenicity of the antigens. The work demonstrates that peptide-based antigens displayed in high densities can induce neutralizing antibody production unlike free peptide. Careful choice of peptides can deliver better candidates. Also, small size allows easy improvisation. Production in bacteria offers cheaper and robust purification option.

High-throughput affinity measurements of direct interactions between activation domains and co-activators.

Sequence-specific activation by transcription factors is essential for gene regulation1,2. Key to this are activation domains, which often fall within disordered regions of transcription factors3,4 and recruit co-activators to initiate transcription5. These interactions are difficult to characterize via most experimental techniques because they are typically weak and transient6,7. Consequently, we know very little about whether these interactions are promiscuous or specific, the mechanisms of binding, and how these interactions tune the strength of gene activation. To address these questions, we developed a microfluidic platform for expression and purification of hundreds of activation domains in parallel followed by direct measurement of co-activator binding affinities (STAMMPPING, for Simultaneous Trapping of Affinity Measurements via a Microfluidic Protein-Protein INteraction Generator). By applying STAMMPPING to quantify direct interactions between eight co-activators and 204 human activation domains (>1,500 K ds), we provide the first quantitative map of these interactions and reveal 334 novel binding pairs. We find that the metazoan-specific co-activator P300 directly binds >100 activation domains, potentially explaining its widespread recruitment across the genome to influence transcriptional activation. Despite sharing similar molecular properties (e.g. enrichment of negative and hydrophobic residues), activation domains utilize distinct biophysical properties to recruit certain co-activator domains. Co-activator domain affinity and occupancy are well-predicted by analytical models that account for multivalency, and in vitro affinities quantitatively predict activation in cells with an ultrasensitive response. Not only do our results demonstrate the ability to measure affinities between even weak protein-protein interactions in high throughput, but they also provide a necessary resource of over 1,500 activation domain/co-activator affinities which lays the foundation for understanding the molecular basis of transcriptional activation.

(Keynote) Automated / Autonomous Platforms for Synthesis, Purification, and Characterization of Quantum Dots

Controlling composition, structure, and thus properties of nanomaterials continues to be of importance to numerous fields. For example, applications of quantum dots range from displays and bioimaging, to metal nanoparticles for (electro-) catalytic conversions. The vastness of synthesis parameter space, in combination with still limited understanding of the nucleation and growth mechanisms for many quantum dots systems still hampers progress in identifying nanomaterials with desired properties for existing and newly envisioned purposes. Furthermore, the identification of optimal synthesis recipes for these nanostructures remains a major hurdle.This presentation will report on the development and application a number of enabling capabilities for the synthesis and characterization of quantum dots, achieved through reactor engineering. We demonstrate how the use of automated flow reactors not only helps in run-to-run reproducibility but also in uncovering mechanistic information. Examples include reactors with dedicated nucleation, growth, and shell formation zones, and investigation that revealed mechanistic insight of how water concentration affects QD synthesis outcome. We also employed an autonomous flow reactor system to map synthesis parameter space of specific QD systems, a project that relied on fast, fully automated in-situ characterization using UV-vis or photo-luminescence in combination with multi-step machine learning workflows The utility of flow reactors, however, is limited when considering chemistries with longer reaction times. Hence, more recently we developed a fully automated batch reactor for parameter space mapping of QD synthesis via hot injection, the most frequently used method in both QD research and production at scale. This batch platform is being augmented with purification capabilities, to address some of the challenges of in-line, in-situ characterization of raw reaction mixtures, and to enable using advanced optical and structural characterization methods to provide insight into the actual structure of the QD materials.

PCN-222/H3PW12O40/graphene oxide@cotton cloth with improved photocatalytic/photothermal properties for water decontamination

In this study, a novel visible-light-driven PCN-222/H3PW12O40/graphene oxide@cotton cloth (PCN-222/HPW/GO@Cot) photocatalytic platform was developed for efficient water sterilization and purification. PCN-222/HPW/GO ternary photocatalyst was prepared via one-pot solvothermal method, and anchored onto cotton cloth though catechol-amine reaction. By introducing HPW as the quasi-semiconductor co-catalyst and efficient electron trap, the photocatalytic activity of PCN-222 was boosted. Besides, the addition of GO further enhanced electron transfer capability and photothermal properties. PCN-222/HPW/GO@Cot exhibited significantly improved organic dye photodegradation and sterilization efficiencies compared to PCN-222 and PCN-222/HPW coated cotton cloths. It also maintained consistent organic dye removal efficiency over reuse cycles and retained high sterilization activity after repeated washings, indicating its remarkable durability and coating stability. The band positions and the enhanced photocatalytic/photothermal mechanisms were investigated in depth. This research suggests the great potential of PCN-222/HPW/GO@Cot photocatalytic platform for practical water purification and sterilization applications, and the potential to become multifunctional textiles.

Magnetic Nanoparticle-Based Microfluidic Platform for Automated Enrichment of High-Purity Extracellular Vesicles.

Extracellular vesicles (EVs) are available in various biological fluids and have highly heterogeneous sizes, origins, contents, and functions. Rapid enrichment of high-purity EVs remains crucial for enhancing research on EVs in tumors. In this work, we present a magnetic nanoparticle-based microfluidic platform (ExoCPR) for on-chip isolation, purification, and mild recovery of EVs from cell culture supernatant and plasma within 29 min. The ExoCPR chip integrates bubble-driven micromixers and immiscible filtration assisted by surface tension (IFAST) technology. The bubble-driven micromixer enhances the mixing between immunomagnetic beads and EVs, eliminating the need for manual pipetting or off-chip oscillatory incubation. The high-purity EVs were obtained after passing through the immiscible phase interface where hydrophilic or hydrophobic impurities nonspecifically bound to SIMI were removed. The ExoCPR chip had a capture efficiency of 75.8% and a release efficiency of 62.7% for model EVs. We also demonstrated the powerful performance of the ExoCPR in isolating EVs from biological samples (>90% purity). This chip was further employed in clinical plasma samples and showed that the number of GPC3-positive EVs isolated from hepatocellular carcinoma patients was significantly higher than that of healthy individuals. This ExoCPR chip may provide a promising tool for EV-based liquid biopsy and other fundamental research.

Efficient solar-driven interfacial evaporation using conjugated microporous polymer networks synthesized from Buchwald–Hartwig coupling

Solar-driven interfacial evaporation (SDIE) has been considered as an effective and sustainable technology to purify water. To optimize heat localization and regulate water-existing forms via developing excellent photothermal materials is a key strategy to enhance SDIE performance. Herein, three novel conjugated microporous polymer networks (namely PBPs) were readily synthesized via Buchwald-Hartwig coupling between aryl bromides and p-phenylenediamine. Under 1 sun irradiation, the pure PBP-He without any additives delivered an excellent water evaporation of 1.18 and 0.96 kg · m−2 · h−1 using pure water, nature seawater as water source, respectively, and the corresponding energy conversion efficiency was up to 81.1 % and 65.7 %. An ever-enhanced SDIE performance was found in the PBPs networks with increased amine-linked sites density. The study provided a promising CMPs-based photothermal conversion platform for water purification, photothermal therapy, and photothermal sterilization, etc.

Abstract 3190: Twin Fc-immune cell engager (ICE): A novel antibody platform for cancer immunotherapy with enhanced avidity for Fcγ receptors and complement

Abstract Immunoglobulin G (IgG)-based mAbs have become a dominant class of cancer immunotherapy due to their excellent drug-like properties, long half-life, and generally safe profiles. Typical IgG-based therapeutic mAbs have strong binding to target antigens owing to the valency effect of two antigen-binding fragments (Fabs). However, the single crystallizable fragment (Fc) lacks the valency effect to engage immune effector elements. As Fcγ receptors (FcγR) such as FcγRIIa and FcγRIIIa and C1q complement component have low affinity for Fc, IgG-based mAbs require high antibody opsonization density to exhibit avidity for these low affinity Fcγ receptors and C1q. This may also account for the suboptimal efficacy of IgG-based mAbs against tumors with less abundant antigen. To enhance effector functions of IgG1 by overcoming such limitation, we devised a novel platform, Twin Fc-ICE, in which a single Fab region is attached by two Fc regions which allow for high avidity binding to Fcγ receptors and C1q. The two Fc regions of Twin Fc-ICE are arranged in parallel, not in tandem, to adopt natural topologies of multiple Fc regions presented by IgG1-opsonized targets. Supporting the avidity effect of Twin Fc-ICE for FcγR binding, we showed that CTN001, a Twin Fc-ICE with trastuzumab Fab, have markedly increased binding to FcγRs including FcγRI, FcγRIIa and FcγRIIIa compared with trastuzumab. The avid FcγR binding was further demonstrated by flow cytometry analysis of human blood which revealed significantly increased binding of CTN001 relative to trastuzumab for immune cells expressing activating FcγRs such as natural killer (NK) cells, monocytes, and dendritic cells (DCs). The monovalent antigen binding of Twin Fc-ICE requires Fab with high affinity. However, provided that high affinity is achieved, the monovalent Fab adds to the two Fc regions to maximize the Fc load on cancer cells expressing low levels of antigen. Indeed, flow cytometry analysis revealed that treatment of CTN001 alone results in higher Fc load on HER2-expressing cancer cells than combination of two anti-HER2 IgG1 mAbs, trastuzumab and pertuzumab. In consistent with increased Fc load on target cells and avid FcγR binding, CTN001 induces enhanced effector functions such as CDC, ADCC and ADCP against HER2-expressing cancer cells compared with trastuzumab, which will be presented in a separate poster. CTN001 exhibits comparable thermal stability with trastuzumab and can be readily purified to >95% purity with typical purification platforms. The serum half-life of CTN001 at 10 mg/kg intravenous dose in rats is found non-inferior to that of trastuzumab (~12 days versus ~7 days). Taken together, Twin Fc-ICE provides an optimized platform with drug-like properties, which leverages avidity effects of Fc to potentiate therapeutic efficacy of antibody. Citation Format: Eun Shik Choi, Hyunkyu Park, Jiseon Bae, Ki-June Lee, Eunmi Kim, Sujin Yoon, Deokjae Lee, Gi-Hyeok Yang. Twin Fc-immune cell engager (ICE): A novel antibody platform for cancer immunotherapy with enhanced avidity for Fcγ receptors and complement [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3190.

Novel purification platform based on multimodal preparative scale separation of mAb fragments and aggregates

Monoclonal antibodies (mAbs) continue to dominate the biopharmaceutical industry. Certain mAbs are prone to fragmentation and clipping and in these cases, adequate removal of these species is critical during manufacturing. Fragments can be generated during fermentation, purification, storage, formulation, and administration. Their addition to the acidic charge-variant of the purified mAb has been reported to decrease stability and potency of the final product. However, contrary to mAb aggregation, manufacturers have not given much attention to removal of fragments and clipped species and as a result most conventional mAb platforms offer at best limited capabilities for their removal. In this study, we propose a novel purification platform that uses multimodal chromatography and achieves complete removal of a range of mAb fragments and clipped products (25–120 kDa). The utility of the platform has been successfully demonstrated for 2 IgG1s and 2 IgG4s. Further, adequate removal of the various host cell impurities such as host cell proteins (<10 ppm) and host cell DNA (<5 ppb) has been achieved. Finally, the platform was able to deliver adequate removal of high molecular weight impurities (<1 %) and a 30 % clearance of the acidic charge variant. The proposed single step has been shown to deliver what the polishing chromatography and intermediate purification chromatography steps deliver in a traditional mAb platform.

Heparin-affinity chromatography is a generic purification platform for chimeric gag VLPs displaying different viral surface antigens

Chimeric virus-like particles formed by the co-expression of HIV-1 gag capsid protein and surface proteins from other viruses can be used as a platform for production of a wide range of viral vaccines. These virus-like particles can be rapidly produced in an insect cell culture using the baculovirus expression vector system. However, a variety of different process-related impurities such as host cell proteins, double stranded DNA, chromatin, and structurally similar bionanoparticles like extracellular vesicles and baculovirus particles are present in the harvested supernatant and complicate the chromatographic purification steps. Heparan sulfate is the first entry point of a lot of enveloped viruses into the cell. Taking advantage of this functional property, its sub-variant, the glycosaminoglycan heparin immobilized on a chromatography media is a universal tool for the separation of enveloped virus-like particles. A purification process was developed consisting of clarification of the cell culture supernatant by membrane filtration, a pre-purification step using core–shell bead flow-through chromatography with Capto™ Core 700, followed by heparin affinity chromatography with Capto™ Heparin. The combination of Capto™ Core 700 and Capto™ Heparin allows for direct load of the flow-through material from the first into the second step without buffer exchange in between. In addition, lengthy buffer screening, as required for ion exchange and multimodal chromatography, can be omitted. Linear salt gradient elution in heparin affinity chromatography enabled separation of virus-like particles and baculovirus. The platformability of the process was demonstrated by purification of virus-like particles expressing influenza A virus hemagglutinin or neuraminidase, or SARS-CoV-2 spike protein as surface antigens and the virus-like particle without co-expressed surface protein. Western blotting, host cell protein and DNA assays, TCID50 (for residual baculovirus), mass spectrometry, multi-angle light scattering and nanoparticle tracking analysis were used to characterize the different fractions obtained during the purification. A similar elution profile was obtained for all tested virus-like particles and deviations could be explained by batch-to-batch variations and differences in productivity for the different constructs. Double stranded DNA was reduced to 7 – 36 ng/109 particles and host cell protein content was below the limit of quantification; in the main product fraction an average seven-fold reduction of baculovirus and a yield of 9 % − 14 % of bound particles with a concentration of 1.4 – 1.8 × 1010 particles/mL in the eluate were achieved meeting requirements for a viral vaccine. Accordingly, this process is suitable as a downstream process platform for chimeric HIV-1 gag-based VLP products.

SWATH-MS as a strategy for CHO host cell protein identification and quantification supporting the characterization of mAb purification platforms

Host cell proteins (HCPs) are process-related impurities expressed by the host cells during biotherapeutics’ manufacturing, such as monoclonal antibodies (mAbs). Some challenging HCPs evade clearance during the downstream processing and can be co-purified with the molecule of interest, which may impact product stability, efficacy, and safety. Therefore, HCP content is a critical quality attribute to monitor and quantify across the bioprocess.Here we explored a mass spectrometry (MS)-based proteomics tool, the sequential window acquisition of all theoretical fragment-ion spectra (SWATH) strategy, as an orthogonal method to traditional ELISA. The SWATH workflow was applied for high-throughput individual HCP identification and quantification, supporting characterization of a mAb purification platform. The design space of HCP clearance of two polishing resins was evaluated through a design of experiment study. Absolute quantification of high-risk HCPs was achieved (reaching 1.8 and 4.2 ppm limits of quantification, for HCP A and B respectively) using HCP-specific synthetic heavy labeled peptide calibration curves. Profiling of other HCPs was also possible using an average calibration curve (using labeled peptides from different HCPs). The SWATH approach is a powerful tool for HCP assessment during bioprocess development enabling simultaneous monitoring and quantification of different individual HCPs and improving process understanding of their clearance.

Improved mRNA affinity chromatography binding capacity and throughput using an oligo-dT immobilized electrospun polymer nanofiber adsorbent

Increased demand for mRNA-based therapeutics and improved in vitro transcription (IVT) yields have challenged the mRNA purification platform. Hybridization-affinity chromatography with an immobilized oligo-deoxythymidilic acid (oligodT) ligand is often used to capture mRNA through base pairing with the polyadenylated tail. Commercially available oligodT matrices include perfusive cross-linked poly(styrene-divinylbenzene) 50 µm POROS™ chromatography resin beads and convective polymethacrylate CIMmultus® monolithic columns consisting of 2 µm interconnected channels. POROS™ columns may be limited by poor mass transfer for larger mRNAs and slow flowrates, while monoliths can operate at higher flowrates but are limited by modest binding capacity. To enable both high flowrates and binding capacity for mRNA of all lengths, prototype chromatography media was developed by Cytiva using oligodT immobilized electrospun cellulose nanofibers (Fibro™) with a 0.3–0.4 µm pore size. In this work, four polyadenylated mRNAs ranging from ∼1900–4300 nucleotides were used to compare the dynamic binding capacity (DBC) of Fibro™, POROS® and CIMmultus® columns as a function of residence time and binding buffer compositions. Fibro™ improved the DBC ∼2–4-fold higher than CIMmultus® and ∼2–13-fold higher than POROS™ across all residence times, mRNA length, and binding matrix compositions tested. CIMmultus® DBC was least dependent on residence time and mRNA size, while both Fibro™ and POROS™ DBC increased at slower flowrates and with shorter mRNA. Surprisingly, inverse size exclusion (ISE) experiments showed that POROS™ was not limited by diffusion and POROS™ along with CIMmultus® demonstrate higher mRNA permeation however the Fibro™ prototype is not in the final configuration. Lastly, IVT reaction products were subjected to purification and oligodT elution product yield, quality, and purity were consistent across the three matrices investigated. These results highlight the benefits of high DBC and equivalent product profiles offered by the oligodT Fibro™ prototype compared to commercially available oligodT media.

Enhanced oxidation of organic pollutants by regulating the interior reaction region of reactive electrochemical membranes

Reactive electrochemical membrane (REM) emerges as an attractive strategy for the elimination of refractory organic pollutants that exist in wastewater. However, the limited reaction sites in traditional REMs greatly hinder its practical application. Herein, a feed-through coating methodology was developed to realize the uniform loading of SnO2-Sb catalysts on the interior surface of a REM. The uniformly coated REM (Unif-REM) exhibited 2.4 times higher reaction kinetics (0.29 min–1) than that of surface coated REM (Surf-REM) for the degradation of 2 mM 4-chlorophenol (4-CP), rendering an energy consumption as low as 0.016 kWh gTOC–1. The fast degradation of various emerging contaminants, e.g., sulfamethoxazole (SMX), ofloxacin (OFLX), and tetracycline (TC), also confirms its superior oxidation capability. Besides, the Unif-REM exhibited good performance in generating hydroxyl radicals (•OH) and a relatively long service lifetime. The simulation of spatial current distribution demonstrates that the interior reaction region in the Unif-REM channels can be drastically extended, thereby maximizing the surface coupling of mass diffusion and electron transfer. This study offers an in-depth look at the spatially confined reactions in REM and provides a reference for the design of electrochemical systems with economically efficient water purification.

Magnetic-based adsorbing platforms for water and wastewater purification

Magnetic-based adsorbing platforms for water and wastewater purification

Defect-engineered graphitic carbon nitride on carbon cloth supports for the photoelectrocatalytic degradation of organophosphate pesticides

Defect engineering of graphitic carbon nitride photoelectrocatalysts promotes the degradation of fenitrothion (FNT), a persistent organophosphate pesticide, paving the way to the development of eco-friendly platforms for wastewater purification.

Impact of drug-linker on method selection for analytical characterization and purification of antibody-drug conjugates.

In addition to traditional characterisation methods of hydrophobic interaction (HIC) and reverse phase (RP) chromatography, an anion exchange chromatography (AIEX) was developed to analyse and purify antibody drug conjugates (ADCs). Since different drug antibody ratio (DAR) species may impact biological activity, therapeutic index, PK parameters or even potential immunogenicity, homogenous ADC DAR demands have been significantly increasing. To accelerate linker designs, drug screening and ADC DAR purification for in vitro and in vivo studies, we built the analytical toolbox including HIC, RP, AIEX, icIEF, SEC, and MS for downstream ADC DAR purification using HIC and AIEX. The established analytical methods can quickly assess the quality of ADC DAR profiles and provide important information to select the proper ADC DAR purification method. Since drug-linker structures can significantly affect ADC physicochemical properties, and highly impact on selections of analytical methods, we applied both HIC and AIEX characterisation and purification platforms to achieve ADC DAR homogenous. Our experiments also implied that unlike HIC, AIEX could be used to separate DAR4 positional isomers.

Disease-Associated Mutations in Tau Encode for Changes in Aggregate Structure Conformation.

The accumulation of tau fibrils is associated with neurodegenerative diseases, which are collectively termed tauopathies. Cryo-EM studies have shown that the packed fibril core of tau adopts distinct structures in different tauopathies, such as Alzheimer's disease, corticobasal degeneration, and progressive supranuclear palsy. A subset of tauopathies are linked to missense mutations in the tau protein, but it is not clear whether these mutations impact the structure of tau fibrils. To answer this question, we developed a high-throughput protein purification platform and purified a panel of 37 tau variants using the full-length 0N4R splice isoform. Each of these variants was used to create fibrils in vitro, and their relative structures were studied using a high-throughput protease sensitivity platform. We find that a subset of the disease-associated mutations form fibrils that resemble wild-type tau, while others are strikingly different. The impact of mutations on tau structure was not clearly associated with either the location of the mutation or the relative kinetics of fibril assembly, suggesting that tau mutations alter the packed core structures through a complex molecular mechanism. Together, these studies show that single-point mutations can impact the assembly of tau into fibrils, providing insight into its association with pathology and disease.

SPinDP: A High-Speed Distributed Processing Platform for Sampling and Filtering Data Streams

Recently, there has been an explosive generation of streaming data in various fields such as IoT and network attack detection, medical data monitoring, and financial trend analysis. These domains require precise and rapid analysis capabilities by minimizing noise from continuously generated raw data. In this paper, we propose SPinDP (Stream Purifier in Distributed Platform), an open source-based high-speed stream purification platform, to support real-time stream purification. SPinDP consists of four major components, Data Stream Processing Engine, Purification Library, Plan Manager, and Shared Storage, and operates based on open-source systems including Apache Storm and Apache Kafka. In these components, stream processing throughput and latency are critical performance metrics, and SPinDP significantly enhances distributed processing performance by utilizing the ultra-high-speed network RDMA (Remote Direct Memory Access). For the performance evaluation, we use a distributed cluster environment consisting of nine nodes, and we show that SPinDP’s stream processing throughput is more than 28 times higher than that of the existing Ethernet environment. SPinDP also significantly reduces the processing latency by more than 2473 times on average. These results indicate that the proposed SPinDP is an excellent integrated platform that can efficiently purify high-speed and large-scale streams through RDMA-based distributed processing.

Visible-light-driven Au/PCN-224/Cu(II) modified fabric with enhanced photocatalytic antibacterial and degradation activity and mechanism insight

The development of novel photocatalytic platform for efficient water purification and sterilization is of utmost importance due to the significant threat of chronic diseases resulting from water pollution and bacterial contamination. Herein, a high-efficiency, easy-recyclable, and environment-friendly photocatalytic fabric was developed as a water disinfection and organic pollutant degradation platform. In which, microamounts of Au nanoparticles (2 wt%) and Cu(II) clusters (2.5 wt%) co-decorated porphyrinic MOF (Au/PCN-224/Cu(II)) coating was applied as photocatalytic surface, and flexible cotton fabric served as stable support. Au/PCN-224/Cu(II) composite photocatalyst was prepared via a facile photo-deposition procedure followed by impregnation method, and further immobilized onto polydopamine (PDA) pre-treated cotton fabric through catechol-amine reaction. The fabricated photocatalytic platform exhibited excellent ability to generate reactive oxygen species (ROS) by multiple routes under low-power visible light irradiation, and achieved efficient photodisinfection effects for causative organism including Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus) without the obvious leakage of metal ions from the potocatalyst during the reaction. Additionally, the reaction rate constant for organic dye MB photodegradation achieved by Au/PCN-224/Cu(II)@Fabric was 3.73 times higher than that of PCN-224@Fabric, and the degradation intermediates analysis was also conducted. The band alignments of PCN-224, Au/PCN-224, Au/PCN-224/Cu(II) photocatalysts were determined, and the charge transfer mechanism within Au/PCN-224/Cu(II) photocatalyst was elucidated through the utilization of DRS, Mott − Schottky, ESR, EIS, and photocurrent analyses. It is suggested that the deposition of Au and doping of Cu(II) both with small amounts enhanced the light absorption capacity, and also improved the charge separation ability through the surface plasmon resonance (SPR) effect and interfacial electron transfer (IFCT) mechanism. Moreover, Au SPR and Cu doping exhibited a substantial enhancement in photothermal conversion effect, thereby synergistically reinforcing the sterilization performance. This work provides an approach for the development of effective photocatalytic platform based on porphyrinic MOFs combined with trace metal co-catalysts in the application of water disinfection and purification.