Hollow Fiber Filter Research Articles

Scalable isolation of surface-engineered extracellular vesicles and separation of free proteins via tangential flow filtration and size exclusion chromatography (TFF-SEC)

BackgroundExtracellular vesicles (EVs) represent small lipid bilayer structures pivotal in mediating intercellular communication via biomolecular transfer. Their inherent characteristics, including packaging, non-immunogenicity, and biofluid stability, position EVs as promising drug delivery vectors. However, developing clinical quality EVs requires multifaceted technological advancement.MethodsIn this study, a method is introduced for engineering extracellular vesicles (eEVs) from cultured cells and their subsequent isolation using lab-scale tangential flow filtration (TFF). This is the first study to evaluate DNA loading efficacy into EVs isolated by TFF, marking a significant milestone in the field of targeted drug delivery. Initially, cells are transfected with EV-display constructs to facilitate the secretion of eEVs bearing the desired coding molecules. Following brief centrifugation, the cell culture media undergoes filtration using hollow fiber filters. TFF, by applying a constant flow, effectively segregates molecules based on designated molecular weight cut-off (MWCO), enriching particles between 50 and 650 nm.ResultsCompared to conventional methods like ultracentrifugation, TFF demonstrates higher efficiency in removing undesired molecules/aggregates while exerting less stress on EVs. Characterization of eEVs through various assays confirms TFF’s superiority in isolating pure EV populations. Additionally, the necessity of size-exclusion chromatography (SEC) after tangential flow filtration (TFF) becomes evident for effectively removing unbound protein contaminants.ConclusionIn conclusion, TFF-SEC emerges as a scalable and superior approach for eEV isolation, promising significant advancements in clinical applications.

Improved sieving coefficient in perfusion cell culture with reduced effective filtration length of hollow fibers.

The hollow fiber filter is the primary cell-retention device used in high-density perfusion cell culture and often used in an alternating tangential flow (ATF) configuration. The limited commercially available diaphragm pumps for ATF prevent utilization of vertical space when scaling beyond 500 L. Stacking hollow fiber filters coupled with viscous cell culture imposes vacuum pressure exceeding facility capabilities. Additionally, the longer filter assembly increases the hold-up volume and exceeds the diaphragm pump's fluid exchange capacity. The conventional tangential flow filtration (TFF) configuration circumvents this issue by exchanging culture from the bioreactor and cell-retention device in a unidirectional recirculation loop; however, the increased filter length when scaled up exacerbates the TFF's inherent issue with product retention from Starling flow. Stacking commercially available 20 cm TFF filters to make up the similar single-module length TFF used for the platform 3 and 50 L perfusion process at 41.5 and 65 cm, respectively, attempts to reduce fouling caused by Starling flow. The permeate of a single-module filter is partitioned into short independent segments through serially stacked filters, each harvested separately. By partitioning the permeate, the sieving coefficient increased for both 3 and 50 L scales. Reduction of Starling flow was confirmed with lower total hydraulic membrane resistance throughout the culture. This work demonstrates a method for increasing sieving coefficient and filter capacity by stacking TFF filters with independent permeate streams.

Off-line chemical cleaning reduces the spatial heterogeneity of membrane fouling in full-scale membrane bioreactors: A case study

Full-scale membrane bioreactors (MBRs) are widely employed for wastewater treatment. Nevertheless, the spatial heterogeneity of membrane fouling within full-scale MBR modules has received limited attention, especially regarding the impact of off-line chemical cleaning on this spatial heterogeneity. This study, based on 192 hollow fiber samples collected from two full-scale MBRs, investigated the spatial fouling heterogeneity before and after off-line chemical cleaning. The membrane samples were categorized into different groups based on sampling locations, and statistical tools were employed to analyze spatial fouling heterogeneity. The results of foulant mass and filtration resistance of samples confirmed the presence of spatial fouling heterogeneity within the MBR modules after 5 months of operation. The off-line chemical cleaning removed organic foulants more effectively than inorganic foulants, and interestingly, it effectively reduced the spatial fouling heterogeneity. Computational fluid dynamics analysis in the cleaning tank and the chlorine concentration distribution indicated that chemical cleaning occurred without mass transfer or reaction limitations. We further offer insights into the reduction of spatial fouling heterogeneity by clarifying the fouling layer structure and the role of off-line chemical cleaning. Our findings suggest that with an appropriate cleaning strategy, the hollow fiber filtration performance can be rebalanced before a new operation cycle.

Contributions of Chinese hamster ovary cell derived extracellular vesicles and other cellular materials to hollow fiber filter fouling during perfusion manufacturing of monoclonal antibodies.

Hollow fiber filter fouling is a common issue plaguing perfusion production process for biologics therapeutics, but the nature of filter foulant has been elusive. Here we studied cell culture materials especially Chinese hamster ovary (CHO) cell-derived extracellular vesicles in perfusion process to determine their role in filter fouling. We found that the decrease of CHO-derived small extracellular vesicles (sEVs) with 50-200 nm in diameter in perfusion permeates always preceded the increase in transmembrane pressure (TMP) and subsequent decrease in product sieving, suggesting that sEVs might have been retained inside filters and contributed to filter fouling. Using scanning electron microscopy and helium ion microscopy, we found sEV-like structures in pores and on foulant patches of hollow fiber tangential flow filtration filter (HF-TFF) membranes. We also observed that the Day 28 TMP of perfusion culture correlated positively with the percentage of foulant patch areas. In addition, energy dispersive X-ray spectroscopy-based elemental mapping microscopy and spectroscopy analysis suggests that foulant patches had enriched cellular materials but not antifoam. Fluorescent staining results further indicate that these cellular materials could be DNA, proteins, and even adherent CHO cells. Lastly, in a small-scale HF-TFF model, addition of CHO-specific sEVs in CHO culture simulated filter fouling behaviors in a concentration-dependent manner. Based on these results, we proposed a mechanism of HF-TFF fouling, in which filter pore constriction by CHO sEVs is followed by cake formation of cellular materials on filter membrane.

Effect of inner diameter, filter length, and pore size on hollow fiber filter fouling during perfusion cell culture.

As the need for higher volumetric productivity in biomanufacturing grows, biopharmaceutical companies are increasingly investing in a perfusion cell culture process, most commonly one that uses a hollow fiber filter as the cell retention device. A current challenge with using hollow fiber filters is fouling of the membrane, which reduces product sieving and can increase transmembrane pressure (TMP) past process limitations. In this work, the impact of hollow fiber filter geometries on product sieving and hydraulic membrane resistance profiles is evaluated in a tangential flow filtration (TFF) perfusion system. The hollow fibers tested had lengths ranging from 19.8 to 41.5 cm, inner diameters (IDs) ranging from 1.0 to 2.6 mm, and pore sizes of 0.2 or 0.65 μm. The results showed that the shortest hollow fibers experienced higher product sieving while larger IDs contributed to both higher product sieving and lower hydraulic membrane resistances, illustrating the impact of filter geometry on process performance. The results also showed 0.2 μm pore size filters maintain higher product sieving, but also higher membrane resistances compared to 0.65 μm pore size filters. This study highlights the need for optimized hollow fiber filter geometries to maximize use of the membrane area, which in turn can reduce production costs and increase scalability of the perfusion process.

Selective ion transport in large-area graphene oxide membrane filters driven by the ionic radius and electrostatic interactions.

Filters made of graphene oxide (GO) are promising for purification of water and selective sieving of specific ions; while some results indicate the ionic radius as the discriminating factor in the sieving efficiency, the exact mechanism of sieving is still under debate. Furthermore, most of the reported GO filters are planar coatings with a simple geometry and an area much smaller than commercial water filters. Here, we show selective transport of different ions across GO coatings deposited on standard hollow fiber filters with an area >10 times larger than typical filters reported. Thanks to the fabrication procedure, we obtained a uniform coating on such complex geometry with no cracks or holes. Monovalent ions like Na+ and K+ can be transported through these filters by applying a low electric voltage, while divalent ions are blocked. By combining transport and adsorption measurements with molecular dynamics simulations and spectroscopic characterization, we unravel the ion sieving mechanism and demonstrate that it is mainly due to the interactions of the ions with the carboxylate groups present on the GO surface at neutral pH.

Efficacy and Safety of Modified Polyethersulfone Hemodialysis Filters in Comparison to Conventional Filters: A Non-inferiority Clinical Trial

Background: The efficacy and quality of hemodialysis (HD) are closely related to the dialyzer characteristics. Objectives: This study aimed to determine the efficacy and complications of modified polyethersulfone (m-PES) -1.5 hollow fiber filters in comparison to conventional filters during HD. Methods: This non-inferiority crossover randomized clinical trial was performed in adult dialysis units at three HD centers within May 2019 to March 2020. The patients were randomly enrolled in two groups. Group A was first put on HD for six sessions with a low or high flux smart flux filter (m-PES-1.5 hollow fiber), which was made in Italy by Medica S.P.A. Group. Group B was hemodialyzed with a corresponding low or high flux filter made in Iran by Meditechsys Company. After a two-week clearance phase, the patients were dialyzed for six sessions with the opposite filter of the first six sessions. Laboratory variables, such as blood urea nitrogen and creatinine, were measured. Kt/V (i.e., a measurement of HD efficacy) and urea reduction ratio (URR) were calculated. Additionally, blood pressure was monitored. Results: A total of 40 patients were entered into the final analysis. No matter which filter was used, no statistically significant differences were observed in URR, creatinine, Kt/V, and blood pressure at different times during dialysis between the two types of filters. Packaging problems (P < 0.001) and blood clotting (P = 0.009) were two more frequent complications in the m-PES group. Conclusions: This study showed that smart flux m-PES-1.5 hollow fiber filters are similar to Meditechsys Company filters.

Handheld virus concentration method using a hollow fiber filter module

A virus concentration method is required for viral vaccine manufacturing and virus-related research. However, concentration methods, such as ultracentrifugation, often require capital investment. We report a simple and easy-to-use handheld syringe method for virus concentration using a hollow fiber (HF) filter module, which can be applicable to viruses of different sizes, without incorporating any special machines or reagents. This virus concentration method does not use pumps, which might cause shear stress for virus particles; therefore, it is useful for stress-sensitive virus particles, and virus-like particles, as well as other proteins.The clarified harvest of flavivirus (Zika virus) was concentrated using an HF filter module and compared with a centrifugal ultrafiltration device (CUD) for demonstration of the HF filter method. The HF filter method achieved concentration of the virus solution in less time than the CUD. The yield comparison of the recovered virus solution indicated that recovery from the developed method was comparable to using the CUD, and infectivity was maintained throughout.•The Zika virus was concentrated from 200 mL to 5 mL within 45 min using the HF filter and handheld syringe module method.•The handheld HF filter method may be applicable to stress-sensitive viruses and proteins of different sizes.•The virus concentration process should be conducted in a safety cabinet, which is preferred for virus containment.

Study forecasts CAGR of 6.5% for hollow-fibre filtration market to 2028

The hollow-fibre filtration market worldwide is set to reach US$3605.5 million by 2028, with a compound annual growth rate (CAGR) of 6.5% during the review period, forecasts a study published by Valuates Reports.

Evaluating filter functionality and user competence after a hollow fiber membrane filter intervention in Liberia

Abstract In Liberia, access to safe water is not universal, and waterborne diseases like diarrhea run rampant. As part of a larger border-to-border clean water project in Liberia, hollow membrane fiber filters were distributed to households in remote and/or small villages across Liberia. While filter efficacy has been demonstrated in the laboratory, studies of filter efficacy in real-world settings yield more mixed results. Intervention efficacy in Liberia was evaluated by assessing (1) user ability to correctly filter and backwash and (2) filter functioning at follow-up visits approximately 2 and 8 weeks post-intervention. Ultimately, the results supported the efficacy of this intervention. At arrival of both follow-ups, over 95% of filters were functioning properly and the majority of issues were resolved during visits. This supported the short-term durability of the filters and the importance of follow-up visits for repairs. Furthermore, the vast majority of households were able to correctly demonstrate filtering and backwashing: 88.47% at the first follow-up and 91.79% at the second. This slight increase may indicate the value of follow-up visits as educational tools. The widescale distribution of point-of-use filters as a mechanism for clean water should include on-going education and affordable filter repair and replacement opportunities.

Treatment of Municipal Wastewater by Hollow Fiber Dynamic Membrane Bioreactors

AbstractIn this study, an aerobic membrane bioreactor (MBR) with a polyester fabric hollow fiber filter is used for municipal wastewater treatment. This polyester fabric filter is used as support material for dynamic membrane formation in the MBR. High treatment performance is obtained in the permeate achieving over 93% of chemical oxygen demand removal efficiency and low particulate material concentration (<10 mg L−1). The system is operated for 58 d without any clogging. The average transmembrane value is found as around 598 mbar after the system reaches stable conditions. Morphological analyses including confocal laser scanning microscopy, environmental scanning electron microscopy, energy‐dispersive X‐ray, and Fourier transform infrared spectroscopy are performed to obtain a detailed understanding of the cake layer formation. The use of hollow fiber dynamic membranes can provide compact membrane volume with high surface area, thus, the volume of the reactor can be further decreased.

Failure of a hollow-fibre shower filter device to prevent exposure of patients to Pseudomonas aeruginosa

Pseudomonas aeruginosa in hospital water is a risk for invasive infection. Point-of-use (POU) filters are used to reduce patient exposure to the organism, and hollow-fibre filters are becoming more popular. However, retrograde colonization of the filter mechanism may contaminate the effluent. To assess the efficacy of POU filter head (polysulfone; hollow-fibre matrix) shower filters in preventing the exposure of high-risk patient groups to P.aeruginosa. Pre-flush (opening the outlet and collecting the first 100mL of water) samples were analysed to measure P.aeruginosa contamination from 25 shower outlets (∼21% of all showers on the six wards), with and without a hollow-fibre filter. P.aeruginosa was measured in a subset of outlets harbouring P.aeruginosa (sampling period 19th August 2019 to 10th January 2020). Water from all 25 showers was heavily colonized [>300colony-forming units (cfu)/mL] with P.aeruginosa at the showerhead. P.aeruginosa was found in 32% (8/25) of post-filter shower water effluent samples with a geometric mean of 4x106cfu/mL (N=4) (6.8x104-2x108). Filters were sampled at 15-150 days of use (median 15 days), with 26% (6/23) of filter units becoming colonized before the expiry date. POU filter showerhead units may not be effective in preventing exposure of vulnerable patients to P.aeruginosa in hospital water due to retrograde contamination (external contamination of the showerhead passed back to the filter cartridge itself) or failure of the hollow-fibre filter matrix. Reliance should not be placed on the use of hollow-fibre filters to protect patients from exposure to P.aeruginosa without repeated microbiological monitoring.

Micro-Bioreactors in Space: Case Study of a Yeast (Saccharomyces cerevisiae) Bioreactor With a Non-Invasive Monitoring Method

Bioreactors in space have applications from basic science to microbial factories. Monitoring bioreactors in microgravity has challenges with respect to fluidics, aeration, sensor size, sample volume and disturbance of medium and cultures. We present a case study of the development of small bioreactors and a non-invasive method to monitor dissolved oxygen, pH, and biomass of yeast cultures. Two different bioreactor configurations were tested for system volumes of 60 ml and 10.5 ml. For both configurations, the PreSens SFR vario, an optical sensor array, collected data autonomously. Oxygen and pH in the cultures were monitored using chemically doped spots, 7 mm in diameter, that were fixed to the bottom of sampling chambers. Spots emitted a fluorescent signal for DO and pH when reacted with oxygen molecules and hydrogen ions, respectively. Biomass was sensed using light reflectance at centered at 605 nm. The, optical array had three light detectors, one for each variable, that returned signals that were pre- and post-calibrated. For heterotrophic cultures requiring oxygen and respiring carbon dioxide, a hollow fiber filter, in-line with the optical array, oxygenated cells and remove carbon dioxide. This provided oxygen levels that were sufficient to maintain aerobic respiration for steady state conditions. Time series of yeast metabolism in the two bioreactors are compared and discussed. The bioreactor configurations can be easily be modified for autotrophic cultures such that carbon dioxide is enhanced and oxygen removed, which would be required for photosynthetic algal cultures.

Membrane aging effects on water recovery during full-scale potable reuse: Mathematical optimization of backwashing frequency for constant-flux microfiltration

One tool in efforts to tackle the ever growing problem of water scarcity is municipal wastewater reclamation to produce drinking water. Microfiltration (MF) is a central technology for potable reuse because it is highly effective in removing pathogenic protozoa, bacteria, and other colloids and for reverse osmosis pretreatment. However, as microfiltered materials accumulate at the membrane surface, its productivity is reduced requiring periodic removal of foulants. A mathematical model of MF is described in the context of hollow fiber filtration that focused on optimizing constant flux operation with backwashing. Design curves were also proposed for determining backwash timing. The model analysis is evaluated against real-world MF fouling for membranes that range in age from a few weeks to three years, observed at the world’s largest water reuse facility operated by the Orange County Water District. The presented model compares well with the full-scale operational data, and model parameters accurately capture variations in fouling kinetics with membrane age, providing clues to changes in optimal regeneration timing and frequency as membrane performance declines over long time scales.

Recovery of SARS-CoV-2 from large volumes of raw wastewater is enhanced with the inuvai R180 system

Wastewater-based epidemiology (WBE) for severe acute respiratory syndrome Coronavirus 2 (SARS-CoV-2) is a powerful tool to complement syndromic surveillance. Although detection of SARS-CoV-2 in raw wastewater may be prompted with good recoveries during periods of high community prevalence, in the early stages of population outbreaks concentration procedures are required to overcome low viral concentrations. Several methods have become available for the recovery of SARS-CoV-2 from raw wastewater, generally involving filtration. However, these methods are limited to small sample volumes, possibly missing the early stages of virus circulation, and restrained applicability across different water matrices. The aim of this study was thus to evaluate the performance of three methods enabling the concentration of SARS-CoV-2 from large volumes of wastewater: i) hollow fiber filtration using the inuvai R180, with an enhanced elution protocol and polyethylene glycol (PEG) precipitation; ii) PEG precipitation; and iii) skimmed milk flocculation. The performance of the three approaches was evaluated in wastewater from multiple wastewater treatment plants (WWTP) with distinct singularities, according to: i) effective volume; ii) percentage of recovery; iii) extraction efficiency; iv) inhibitory effect; and v) the limits of detection and quantification. The inuvai R180 system had the best performance, with detection of spiked control across all samples, with average recovery percentages of 68% for porcine epidemic diarrhea virus (PEDV), with low variability. Mean recoveries for PEG precipitation and skimmed milk flocculation were 9% and 14%, respectively. The inuvai R180 enables the scalability of volumes without negative impact on the costs, time for analysis, and recovery/inhibition. Moreover, hollow fiber ultrafilters favor the concentration of different microbial taxonomic groups. Such combined features make this technology attractive for usage in environmental waters monitoring.

Non-pharmacological interventions for preventing clotting of extracorporeal circuits during continuous renal replacement therapy.

Non-pharmacological interventions for preventing clotting of extracorporeal circuits during continuous renal replacement therapy.

Preparation and electrochemical properties of porous carbon materials derived from waste hollow fiber filter membrane

In this paper, waste hollow fiber filter membrane is used as a carbon source and KOH as an activator under an atmosphere of protective argon gas to prepare hollow fiber filter membrane-based porous carbon material ZKC-T via a high-temperature pyrolysis activation method. Hollow fiber filter membrane-based porous carbon ZKC-T was found to consist of amorphous carbon, with an irregular three-dimensional network interconnecting pore structure, specific surface area of up to 2934 m2 g−1, and a reasonable pore size distribution. Cyclic voltammetry and constant current charge and discharge tests showed that the hollow fiber filter-based porous carbon material ZKC-700 has excellent electrochemical performance. The carbon material shows a specific capacitance of up to 289 F g−1 at a current density of 1 A g−1 in 6 M KOH electrolyte. In addition, the material shows good cycle stability; after 5000 charge and discharge cycles, the capacitance retention rate is as high as 92.8%. The hollow fiber filter membrane-based porous carbon prepared by this method exhibits excellent electrochemical properties.

Mesenchymal stromal cell delivery via an ex vivo bioreactor preclinical test system attenuates clot formation for intravascular application

While mesenchymal stromal cells are an appealing therapeutic option for a range of clinical applications, their potential to induce clotting when used systemically remains a safety concern, particularly in hypercoagulable conditions, such as in patients with severe COVID‐19, trauma, or cancers. Here, we tested a novel preclinical approach aimed at improving the safety of mesenchymal stromal cell (MSC) systemic administration by use of a bioreactor. In this system, MSCs are seeded on the exterior of a hollow‐fiber filter, sequestering them behind a hemocompatible semipermeable membrane with defined pore‐size and permeability to allow for a molecularly defined cross talk between the therapeutic cells and the whole blood environment, including blood cells and signaling molecules. The potential for these bioreactor MSCs to induce clots in coagulable plasma was compared against directly injected “free” MSCs, a model of systemic administration. Our results showed that restricting MSCs exposure to plasma via a bioreactor extends the time necessary for clot formation to occur when compared with “free” MSCs. Measurement of cell surface data indicates the presence of known clot inducing factors, namely tissue factor and phosphatidylserine. Results also showed that recovering cells and flushing the bioreactor prior to use further prolonged clot formation time. Furthermore, application of this technology in two in vivo models did not require additional heparin in fully anticoagulated experimental animals to maintain target activated clotting time levels relative to heparin anticoagulated controls. Taken together the clinical use of bioreactor housed MSCs could offer a novel method to control systemic MSC exposure and prolong clot formation time.

Determination of the efficiency of filtration of cultures from microalgae and bacteria using hollow fiber filters

Sampling with hollow fiber filters can revolutionize sampling routines. Larger volumes, more representative of the population, can be taken and concentrated for downstream analysis to provide a better estimation of the populations being sampled.

Core-shell graphene oxide-polymer hollow fibers as water filters with enhanced performance and selectivity.

Commercial hollow fiber filters for micro- and ultrafiltration are based on size exclusion and do not allow the removal of small molecules such as antibiotics. Here, we demonstrate that a graphene oxide (GO) layer can be firmly immobilized either inside or outside polyethersulfone-polyvinylpyrrolidone hollow fiber (Versatile PES®, hereafter PES) modules and that the resulting core-shell fibers inherits the microfiltration ability of the pristine PES fibers and the adsorption selectivity of GO. GO nanosheets were deposited on the fiber surface by filtration of a GO suspension through a PES cartridge (cut-off 0.1-0.2 μm), then fixed by thermal annealing at 80 °C, rendering the GO coating stably fixed and unsoluble. The filtration cut-off, retention selectivity and efficiency of the resulting inner and outer modified hollow fibers (HF-GO) were tested by performing filtration on water and bovine plasma spiked with bovine serum albumin (BSA, 66 kDa, ≈15 nm size), monodisperse polystyrene nanoparticles (52 nm and 303 nm sizes), with two quinolonic antibiotics (ciprofloxacin and ofloxacin) and rhodamine B (RhB). These tests showed that the microfiltration capability of PES was retained by HF-GO, and in addition the GO coating can capture the molecular contaminants while letting through BSA and smaller polystyrene nanoparticles. Combined XRD, molecular modelling and adsorption experiments show that the separation mechanism does not rely only on physical size exclusion, but involves intercalation of solute molecules between the GO layers.