Virus Removal Research Articles

Evaluation of woodchips-amended biosand filter for nitrate and MS2 bacteriophage reduction.

In this study, two types of woodchip-amended biosand filters (Filter A sand: woodchip = 33%: 67% versus Filter B sand: woodchip = 50%: 50%, by volume) were constructed, and their abilities to remove MS2 bacteriophage and nitrate were investigated. The results indicated that Filter A and Filter B could reduce nitrate up to 40 and 36%, respectively, indicating that the nitrate reduction increased with the increase in woodchip proportion. The study underscores a positive correlation between nitrate reduction and proportional increase in woodchip content, implying the potential for fine-tuning nitrate removal by varying sand-woodchip compositions. W-BSFs could remove MS2 bacteriophage to 1.91-log10 (98.8%) by Filter A and 1.88-log10 (98.7%) by Filter B over 39 weeks. The difference in sand-woodchip proportion did not significantly impact the MS2 reduction, demonstrating that a single W-BSF can maintain its virus removal performance fairly well over a long-term period. These results indicated that the nitrate reduction could be adjusted by varying sand-woodchip contents without impacting virus removal performance. Microbial community analysis indicated that the nitrate removal by the W-BSFs could be attributed to the denitrifying bacteria, such as the family Streptomycetaceae, the genera Pseudomonas, and Bacillus, and relative abundances of the phylum Nitrospirae.

Adapting virus filtration to continuous processing: Effects of product and process variability on filtration performance.

Virus filtration (VF) is an important unit operation in the manufacture of biotherapeutics that provides robust removal of potential virus contaminants. Small virus removal can be impacted by the low operating pressures and potential depressurization events that are often associated with continuous operations where increased operational flexibility for higher loading at low flux and low pressure is required. In this study, we evaluated the impact of low flux (7 LMH) and pressure interruptions on minute virus of mice (MVM) removal. We used long-term filtrations conducted to a target throughput of 1000 L/m2 with four different monoclonal antibodies on small-scale hollow fiber virus filters with a hydrophilic modified polyvinylidene fluoride membrane. These conditions are certainly challenging for any VF operation and ensuring robust viral clearance under such conditions is critical to the design and implementation of continuous VF. Planova BioEX filters effectively removed MVM at 4 log or greater when run continuously for up to 6 days. Interestingly, pressure increases associated with filter fouling over the duration of long-term filtrations were shown to be reflective of load material variability and could be remediated by implementation of an inline prefilter. Pressure interruptions had minimal impact on overall MVM logarithmic reduction value. Effective virus removal was achieved with pressure increases being largely product-specific, which demonstrates the capability of the virus filter to remove virus independent of pressure increases that are expected to occur with increased protein load.

Comparison on removal performance of virus, antibiotic-resistant bacteria, cell-associated and cell-free antibiotic resistance genes, and indicator chemicals by ozone in the filtrated secondary effluent of a sewage treatment plant

Due to the widespread appearance of viruses, antibiotic-resistant bacteria (ARBs), and antibiotic resistance genes (ARGs) in the aquatic environment, more powerful oxidation processes such as ozonation are needed to enhance the efficiency of their inactivation and removal during wastewater treatment. However, information is lacking on the elimination rates of viruses, ARBs, cell-associated ARGs (ca-ARGs), and cell-free ARGs (cf-ARGs) during ozonation. This study examined the kinetics and dose-dependent inactivation of a virus (MS2 coliphage) and an ARB (Ampicillin-resistant [AmpR] E. coli) and the removal of ca- and cf-ARGs (plasmid-encoded blaTEM) by ozonation in a filtered secondary effluent (SE) of a municipal sewage treatment plant (STP). In addition, the ozonation kinetics of carbamazepine (CBZ) and metoprolol (MTP)—ubiquitous organic micropollutants with different removal rate constants—were also investigated in order to monitor their effectiveness as indicators for the abovementioned biological risk factors. Our results showed that ozonation was an efficient way to remove MS2, AmpRE. coli, ARGs, CBZ, and MTP. We investigated the kinetics of their inactivation/removal with respect to exposure in terms of CT (dissolved ozone concentration C and contact time T) value, and found their inactivation/removal constants were in the following order: MS2 (8.66 ×103 M−1s−1) ≈ AmpRE. coli (8.19 ×103 M−1s−1) > cf-ARG (3.95 ×103 M−1s−1) > CBZ (3.21 ×103 M−1s−1) > ca-ARG (2.48×103 M−1s−1) > MTP (8.35 ×102 M−1s−1). In terms of specific ozone dose, > 5-log inactivation of MS2 was observed at > 0.30 mg O3/mg DOC, while > 5-log inactivation of AmpRE. coli was confirmed at 1.61–2.35 mg O3/mg DOC. Moreover, there was almost no removal of ca-ARG when the specific ozone dose was < 0.68 mg O3/mg DOC. However, 2.86–3.42-log removal of ca-ARG was observed at 1.27–1.31 mg O3/mg DOC, while 1.14–1.36-log removal of cf-ARG was confirmed at 3.60–4.30 mg O3/mg DOC. As alternative indicators, > 4-log removal of CBZ was observed at > 1.00 mg O3/mg DOC, while > 2-log removal of MTP was confirmed at > 2.00 mg O3/mg DOC. Thus, it was observed that inactivation of E. coli needs a greater ozone dose to achieve the same level of inactivation of AmpRE. coli; for ARGs, cf-ARG can persist longer than ca-ARG if low dosages of ozone are applied in the filtrated SE, CBZ might act as an indicator with which to monitor the inactivation of viruses and ARBs, while MTP might act as an indicator with which to monitor removal of ARGs. Moreover, cf-ARG cannot be neglected even after ozonation due to the possibility that ca-ARGs can become cf-ARGs during ozonation and be discharged with the final effluent, posing a potential risk to the receiving environment.

Implementation of a Sensitive Method to Assess High Virus Retention Performance of Low-Pressure Reverse Osmosis Process.

Human enteric viruses are important etiological agents of waterborne diseases. Environmental waters are usually contaminated with low virus concentration requiring large concentration factors for effective detection by (RT)-qPCR. Low-pressure reverse osmosis is often used to remove water contaminants, but very few studies focused on the effective virus removal of reverse osmosis treatment with feed concentrations as close as possible to environmental concentrations and principally relied on theoretical virus removal. The very low viral concentrations usually reported in the permeates (i.e. at least 5 log of removal rate) mean that very large volumes of water need to be analysed to have sufficient sensitivity and assess the process efficiency. This study evaluates two methods for the concentration of adenoviruses, enteroviruses and MS2 bacteriophages at different viral concentrations in large (< 200 L) and very large (> 200 L) volumes. The first method is composed of two ultrafiltration membranes with low-molecular weight cut-offs while the second method primarily relies on adsorption and elution phases using electropositive-charged filters. The recovery rates were assessed for both methods. For the ultrafiltration-based protocol, recovery rates were similar for each virus studied: 80% on average at high virus concentrations (106-107 viruses L-1) and 50% at low virus concentrations (103-104 viruses L-1). For the electropositive-charged filter-based method, the average recoveries obtained were about 36% for ADV 41, 57% for CV-B5 and 1.6% for MS2. The ultrafiltration-based method was then used to evaluate the performance of a low-pressure reverse osmosis lab-scale pilot plant. The retentions by reverse osmosis were similar for all studied viruses and the validated recovery rates applied to the system confirmed the reliability of the concentration method. This method was effective in concentrating all three viruses over a wide range of viral concentrations. Moreover, the second concentration method using electropositive-charged filters was studied, allowing the filtration of larger volumes of permeate from a semi-industrial low-pressure reverse osmosis pilot plant. This reference method was used because of the inability of the UF method to filter volumes on the order of one cubic metre.

Impact of proteins and protein fouling on virus retention during virus removal filtration.

Virus filtration is a crucial step in ensuring the high levels of viral clearance required in the production of biotherapeutics produced in mammalian cells or derived from human plasma. Previous studies have reported that virus retention is often reduced in the presence of therapeutic proteins due to membrane fouling; however, the underlying mechanisms controlling this behavior are still not well understood. Experimental studies were performed with a single layer of the commercially available dual-layer PegasusTM SV4 virus removal filter to more easily interpret the experimental results. Bacteriophage ФX174 was used as a model parvovirus, and human immunoglobulin (hIgG) and Bovine Serum Albumin (BSA) were used as model proteins. Data obtained with 5 g/L solutions of hIgG showed more than a 100-fold reduction in virus retention compared to that in the protein-free solution. Similar effects were seen with membranes that were pre-fouled with hIgG and then challenged with ФX174. The experimental data were well-described using an internal polarization model that accounts for virus capture and accumulation within the virus filter, with the hIgG nearly eliminating the irreversible virus capture while also facilitating the release of previously captured virus. These results provide important insights into the performance and validation of virus removal filters in bioprocessing.

Integration of Planova filters in manufacturing processes of biologicals improve the virus safety effectively: A review of publicly available data.

The capacity to remove viruses by Planova filters produced by Asahi Kasei, primarily by small virus-retentive filters, were compiled from data in peer-reviewed publications and, partly, publicly available data from presentations at conferences (Planova workshops). Data from more than 100 publications and presentations at conferences covering Planova filters were assessed. The data were grouped according to the different virus filters regarding mean pore sizes and viruses of different sizes for plasma and cell culture derived products. Planova 15N and 20N filters removed parvoviruses below the limit of detection of viruses in the filtrate in approx. 50% of all studies and mean LRFs (log reduction factors) for viruses detected in the filtrate were above 4, demonstrating effective parvovirus reduction. Parvovirus removal capacity increased for Planova BioEX filters as well as for 2 Planova 20N in series. Large viruses as retroviruses (e.g., HIV and MuLV), herpesviruses, flaviviruses and togaviruses were removed effectively by Planova 15N, 20N and BioEX filters and also by Planova 35N filters. Flow interruption, transmembrane pressure, volume and protein concentration per filter area had had no substantial impact on virus removal capacity at manufacturing specification. In conclusion, the incorporation of Planova filters in manufacturing processes of biologicals remove, depending on the filter pore size, small and large viruses from the feed stream reliably. This virus reduction step with an orthogonal mechanism integrated in the manufacturing processes of biologicals, based primarily on size exclusion of viruses, improves the virus safety of these biopharmaceutical products considerably.

Airborne influenza virus shedding by patients in health care units: Removal mechanisms affecting virus transmission.

In this study, we characterize the distribution of airborne viruses (influenza A/B) in hospital rooms of patients with confirmed infections. Concurrently, we monitored fine particulate matter (PM2.5 & PM10) and several physical parameters including the room air exchange rate, temperature, and relative humidity to identify corresponding correlations with virus transport and removal determinants. The results continue to raise concerns about indoor air quality (IAQ) in healthcare facilities and the potential exposure of patients, staff and visitors to aerosolized viruses as well as elevated indoor PM levels caused by outdoor sources and/or re-suspension of settled particles by indoor activities. The influenza A virus was detected in 42% of 33 monitored rooms, with viruses detectible up to 1.5 m away from the infected patient. Active coughing was a statistically significant variable that contributed to a higher positive rate of virus detection in the collected air samples. Viral load across patient rooms ranged between 222 and 5,760 copies/m3, with a mean of 820 copies/m3. Measured PM2.5 and PM10 levels exceeded IAQ daily exposure guidelines in most monitored rooms. Statistical and numerical analyses showed that dispersion was the dominant viral removal pathway followed by settling. Changes in the relative humidity and the room's temperature were had a significant impact on the viral load removal. In closure, we highlight the need for an integrated approach to control determinants of IAQ in patients' rooms.

Virus filtration using small pore virus filter in downstream processing of biotherapeutic products: The effect of operating pressure

Virus filtration is a robust and effective method to remove potential virus contaminants. Planova 20 N, a virus filter form Asahi Kasei Bioprocess, has been widely used in the manufacturing process of biotherapeutics. Previous studies have shown that parvovirus removal by Planova 20 N can be impacted by low operation pressure and depressurization. Therefore, it is critical to define an operating pressure range for robust virus removal. In this work, the effect of pressure combined with depressurization on virus removal by Planova 20 N was investigated. Our studies showed that effective virus removal can be achieved in the pressure range from 0.7 bar to 1.6 bar. The data also suggest that re-starting with higher pressure after depressurization is highly desirable for large-scale manufacture to mitigate virus leakage risk. In addition, skipping buffer flush post mainstream filtration minimizes the likelihood of depressurization.

Comparison of kits for SARS-CoV-2 extraction in liquid and passive samples.

Effective extraction and detection of viral nucleic acids from sewage are fundamental components of a successful SARS-CoV-2 sewage surveillance programme. As there is no standard method employed in sewage surveillance, understanding the performance of different extraction kits in the recovery of SARS-CoV-2 and the impact that PCR inhibitors have on quantification is essential to minimize data discrepancies caused by sample extraction. Three commercial nucleic acid extraction kits: the RNeasy PowerSoil Total RNA Kit (PS), the RNeasy PowerMicrobiome Kit (PMB), and the MagMAX™ Microbiome Ultra Nucleic Acid Isolation Kit (MM), with minor modifications, were evaluated. Their efficacy in recovering viral ribonucleic acid and removal of PCR inhibitors was assessed using two South Australian wastewater matrices-one from a major metropolitan site and one from a regional centre. Both had SARS-CoV-2 present due to active COVID-19 cases in these communities. Overall, the MM kit had a higher recovery of SARS-CoV-2 from the samples tested, followed by PMB and PS. The PMB kit performance was strongly influenced by the sample matrix when compared to the MM kit. It is recommended to assess the performance of extraction kits using different local wastewater matrices to ensure the accuracy and reliability of monitoring results to avoid false reporting.

Data-driven systematic analysis of waterborne viruses and health risks during the wastewater reclamation process

Waterborne viral epidemics are a major threat to public health. Increasing interest in wastewater reclamation highlights the importance of understanding the health risks associated with potential microbial hazards, particularly for reused water in direct contact with humans. This study focused on identifying viral epidemic patterns in municipal wastewater reused for recreational applications based on long-term, spatially explicit global literature data during 2000–2021, and modelled human health risks from multiple exposure pathways using a well-established quantitative microbial risk assessment methodology. Global median viral loads in municipal wastewater ranged from 7.92 × 104 to 1.4 × 106 GC L−1 in the following ascending order: human adenovirus (HAdV), norovirus (NoV) GII, enterovirus (EV), NoV GI, rotavirus (RV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Following secondary or tertiary wastewater treatment, NoV GI, NoV GII, EV, and RV showed a relatively higher and more stable log reduction value with medians all above 0.8 (84%), whereas SARS-CoV-2 and HAdV showed a relatively lower reduction, with medians ranging from 0.33 (53%) to 0.55 (72%). A subsequent disinfection process effectively enhanced viral removal to over 0.89-log (87%). The predicted event probability of virus-related gastrointestinal illness and acute febrile respiratory illnesses in reclaimed recreational water exceeded the World Health Organization recommended recreational risk benchmark (5% and 1.9%, respectively). Overall, our results provided insights on health risks associated with reusing wastewater for recreational purposes and highlighted the need for establishing a regulatory framework ensuring the safety management of reclaimed waters.

Enteric virus removal by municipal wastewater treatment to achieve requirements for potable reuse

Primary and secondary treatment of municipal wastewater contributes to virus removal upstream of advanced purification to produce water for potable reuse. In this study, virus occurrence by cultivable and molecular methods was measured over a 24-month period in raw wastewater influents and secondary effluents from two municipal wastewater treatment plants that together provide the recycled water source for an advanced water purification facility. Using a rank-paired, covariance-based statistical approach, virus log removal values were determined for four wastewater treatment processes that operate in parallel at the two facilities (two activated sludge processes, trickling filter process, and trickling filter/solids contactor process). The trickling filter process exhibited the lowest observed removal of cultivable enteric virus with a median removal of 1.0 log10 (or 90% removal) and a 5th percentile log removal of 0.73 (or 82%), compared to the greatest removal observed for one of the activated sludge processes (median log removal of 2.4 or 99.6% and 5th percentile of 2.1 or 99.2%). Median log removal observed for male-specific (MS) and somatic (SOM) coliphage was 1.8 (98.6% removal) and 0.5 (70%), respectively, for trickling filter and 2.9 (99.9%) and 2.0 (99%) for activated sludge. Thus, coliphage removal was fairly similar to removal observed for cultivable enteric virus. The cultivable enteric virus 5th percentile removal (0.7) from the trickling filter treatment process was proposed to the state regulator for credit towards state requirements for virus removal related to groundwater augmentation with purified recycled water. Receiving pathogen removal credits for secondary wastewater treatment would allow for an improved margin (safety factor) of credits beyond the minimum required; and in this case may also increase the number of viable future groundwater recharge sites closer to drinking water production wells by reducing the underground travel time otherwise required to obtain sufficient credits.

Size-based analysis of virus removal filter fouling using fractionated protein aggregates.

Fouling by protein aggregates reduces virus removal filter performance. In the present study, we investigated the effects of different-sized protein aggregates on fouling and aggregate retention in order to better understand the fouling mechanisms. Human immunoglobulin G was denatured by heating to produce aggregates of various sizes and then fractionated by size exclusion chromatography into different-sized aggregates with a narrow size distribution. The fractionated aggregates were filtered on Planova 20N, a virus removal filter known for its stable filtration capability. Analysis of flux behavior demonstrated different flux decrease patterns for different-sized aggregates. Observation of aggregate retention by staining revealed that larger aggregates were captured closer to the inner surface of the membrane while smaller aggregates penetrated farther into the membrane. These findings demonstrate that Planova 20N has a gradient structure with decreasing pore size from the inner to the outer surface of the membrane. This structure minimizes fouling and enables stable filtration by protecting the smaller pores located closer to the outer surface from clogging by large aggregates. Applying the predominant clogging models to the present filtrations revealed that clogging behavior transitioned from complete blocking to cake filtration as filtration progressed. In this combination model, after a certain number of pores are blocked by complete blocking, newly arrived aggregates begin to accumulate on previously captured aggregates, generating cake between capture layers within the membrane. Application of the approaches described here will facilitate elucidation of membrane fouling and virus removal mechanisms.

The impact of heating, ventilation, and air conditioning design features on the transmission of viruses, including the 2019 novel coronavirus: A systematic review of filtration.

Historically, viruses have demonstrated airborne transmission. Emerging evidence suggests the novel coronavirus (SARS-CoV-2) that causes COVID-19 also spreads by airborne transmission. This is more likely in indoor environments, particularly with poor ventilation. In the context of airborne transmission, a vital mitigation strategy for the built environment is heating, ventilation, and air conditioning (HVAC) systems. HVAC features could modify virus transmission potential. A systematic review was conducted to identify and synthesize research examining the effectiveness of filters within HVAC systems in reducing virus transmission. A comprehensive search of OVID MEDLINE, Compendex, and Web of Science Core was conducted to January 2021. Two authors were involved in study selection, data extraction, and risk of bias assessments. Study characteristics and results were displayed in evidence tables and findings were synthesized narratively. Twenty-three relevant studies showed that: filtration was associated with decreased transmission; filters removed viruses from the air; increasing filter efficiency (efficiency of particle removal) was associated with decreased transmission, decreased infection risk, and increased viral filtration efficiency (efficiency of virus removal); increasing filter efficiency above MERV 13 was associated with limited benefit in further reduction of virus concentration and infection risk; and filters with the same efficiency rating from different companies showed variable performance. Adapting HVAC systems to mitigate virus transmission requires a multi-factorial approach and filtration is one factor offering demonstrated potential for decreased transmission. For filtration to be effective, proper installation is required. Of note, similarly rated filters from different companies may offer different virus reduction results. While increasing filtration efficiency (i.e., increasing MERV rating or moving from MERV to HEPA) is associated with virus mitigation, there are diminishing returns for filters rated MERV 13 or higher. Although costs increase with filtration efficiency, they are lower than the cost of ventilation options with the equivalent reduction in transmission. Systematic review registration: PROSPERO 2020 CRD42020193968.

The role of intermolecular interactions on monoclonal antibody filtration through virus removal membranes.

The removal of viruses by filtration is a critical unit operation to ensure the overall safety of monoclonal antibody (mAb) products. Many mAbs show very low filtrate flux during virus removal filtration, although there are still significant uncertainties regarding both the mechanisms and antibody properties that determine the filtration behavior. Experiments were performed with three highly purified mAbs through 3 different commercial virus filters (Viresolve ® Pro, Viresolve® NFP, and Pegasus™ SV4) with different pore structures and chemistries. The flux decline observed during mAb filtration was largely reversible, even under conditions where the filtrate flux with the mAb was more than 100-fold smaller than the corresponding buffer flux. The extent of flux decline was highly correlated with the hydrodynamic diameter of the mAb as determined by dynamic light scattering. The mAb with the lowest filtrate flux for all 3 membranes showed the largest attractive intermolecular interactions and the greatest hydrophobicity, with the latter determined by binding to a butyl resin in an analytical hydrophobic interaction chromatography column. These results strongly suggest that the flux behavior is dominated by reversible self-association of the mAbs, providing important insights into the design of more effective virus filtration processes and in the early identification of problematic mAbs / solution conditions. This article is protected by copyright. All rights reserved.

Assessment of novel antiviral filter using pseudo-type SARS-CoV-2 virus in fast air velocity vertical-type wind tunnel

Current evidence suggests that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can remain suspended spread in aerosols for longer period of time under poorly ventilated indoor setting. To minimize spreading, application of antiviral filter to capture infectious aerosols and to inactivate SARS-CoV-2 can be a promising solution. This study aimed to develop a method to assess simultaneously the filtration and removal efficiency of aerosolized pseudo-type SARS-CoV-2 using a vertical-type wind tunnel with relatively high face velocity (1.3 m/s). Comparing with the untreated spunlace non-woven filter, the C-POLAR™ treated filter increased the filtration efficiency from 74.2 ± 11.5% to 97.2 ± 1.7%, with the removal efficiency of 99.4 ± 0.051%. The results provided not only solid evidence to support the effectiveness of the cationic polymeric coated filter in fighting against the SARS-CoV-2 pandemic, but also a method to test viral filtration and removal efficiency under relative fast air velocity and with a safer environment to the operators.

Surface modification of PVDF hollow fiber ultrafiltration membranes for biopharmaceutical products, virus, and bacterial phage removal technology

In this study, radicals were chemically formed at the hydroxyl (-OH) sites on the surface of polyvinylidene fluoride (PVDF) hollow fiber membranes, and anionic-hydrophilic and hydrophobic materials were grafted onto the surface through -OH radical reaction to generate anions (−28, −41 mV). These membranes were then used as biomembranes (BMs) and showed excellent properties. We also studied the effects of chemical surface modification and reduction in pore size (60–130 nm) according to molecular weight. After anionic modification, the electrostatic repulsion between the pore surface and the bacteria prevents adsorption when anionic bacteria pass through the pores. Particles do not pass through the asymmetric pores due to agglomeration, resulting in separation. The hydrophilic membrane had a high initial water flux, but the flux gradually decreased due to fouling. In contrast, the hydrophobic modified membrane had a low initial water flux that eventually stabilized due to excellent fouling resistance. The protein recovery ratio (PRR) was 98.8 % for hydrophilic particles and 99.2 % for hydrophobic molecules, showing a higher PRR than the pristine membrane (96.4 %). An evaluation of removal of the gram-negative bacteria Brevundimonas diminuta (Bd) showed that no bacteria in membrane-purified solutions, confirming 100 % removal efficiencies. The developed membrane can be used in biopharmaceutical production and removal of proteins, retro viruses, and bacteria.

Protein fouling during constant-flux virus filtration: Mechanisms and modeling.

As biomanufacturers consider the transition from batch to continuous processing, it will be necessary to re-examine the design and operating conditions for many downstream processes. For example, the integration of virus removal filtration in continuous biomanufacturing will likely require operation at low and constant filtrate flux instead of the high (constant) transmembrane pressures (TMPs) currently employed in traditional batch processing. The objective of this study was to examine the effect of low operating filtrate flux (5-100 L/m2 /h) on protein fouling during normal flow filtration of human serum Immunoglobulin G (hIgG) through the Viresolve® Pro membrane, including a direct comparison of the fouling behavior during constant-flux and constant-pressure operation. The filter capacity, defined as the volumetric throughput of hIgG solution at which the TMP increased to 30 psi, showed a distinct minimum at intermediate filtrate flux (around 20-30 L/m2 /h). The fouling data were well-described using a previously-developed mechanistic model based on sequential pore blockage and cake filtration, suitably modified for operation at constant flux. Simple analytical expressions for the pressure profiles were developed in the limits of very low and high filtrate flux, enabling rapid estimation of the filter performance and capacity. The model calculations highlight the importance of both the pressure-dependent rate of pore blockage and the compressibility of the protein cake to the fouling behavior. These results provide important insights into the overall impact of constant-flux operation on the protein fouling behavior and filter capacity during virus removal filtration using the Viresolve® Pro membrane.

Virus breakthrough behavior during virus filtration: Effects of virus/protein concentration and operating conditions

Virus filtration in biopharmaceutical downstream process ensures viral safety with removal efficiency greater than 99.99%. Despite its robust performance, it has been reported that viruses can escape through the membrane under certain conditions. This study aimed to investigate virus breakthrough using Viresolve® Pro membrane with PP7 bacteriophage under high bacteriophage titer, high protein concentration, flow interruption, and low-flux operation. The results show high virus removal performance (LRV >6.8) up to 109 PFU/mL for 200 L/m2, with the first phage breakthrough observation over 1012 PFU/m2 of phage challenge in the membrane. As the protein concentration increased in the coexistence condition, a greater number of phages passed through the membrane. During the post-buffer flush and the operation at low filtrate flux, diffusion plays an important role in the escape of phages, which were detected as high as 1012 PFU/m2, but as low as 109 PFU/m2. These results suggest that the virus-retentive membrane may become susceptible to undesired virus breakthrough when exposed to phage challenges of > 1012 PFU/mL, providing important key factors to ensure viral safety during the downstream process of biopharmaceutical production.

Predicting virus filter performance using an advanced membrane structural model

Virus filters are essential in bioprocessing for safe therapeutic protein production. Understanding the correlation between virus filter structure and performance is important for efficient process design. By applying a multilayer structural model comprised of theoretical layers having pore size distributions, to published visualization data, we demonstrate in the present study that virus removal performance and filtration behavior can be quantitatively calculated. Virus filter structure can be classified into asymmetric high porosity, symmetric high density and laminated structure types. All types exhibit sufficient virus removal properties when used in normal processes. Laminated structure filters show extremely high virus particle removal performance, achieving the most robust virus removal even in filtration with process pauses, although flux decay tends to occur during filtration of solutions containing even very small amounts of aggregate. Conversely, asymmetric high porosity filters may show lower virus removal in processes involving multiple pauses that exceed practical conditions but shows stable filtration behavior with lower increase in pressure and lower flux decay even for the filtration of solutions containing aggregates. Such filtration behavior and virus removal properties can be quantitatively expressed and predicted by numerical calculations using a model based on the theoretical multilayer structure.

Membrane modification in enhancement of virus removal: A critical review

Many waterborne diseases are related with viruses, and COVID-19 worldwide has raised the concern of virus security in water into the public horizon. Compared to other conventional water treatment processes, membrane technology can achieve satisfactory virus removal with fewer chemicals, and prevent the outbreaks of viruses to a maximal extent. Researchers developed new modification methods to improve membrane performance. This review focused on the membrane modifications that enhance the performance in virus removal. The characteristics of viruses and their removal by membrane filtration were briefly generalized, and membrane modifications were systematically discussed through different virus removal mechanisms, including size exclusion, hydrophilic and hydrophobic interactions, electronic interactions, and inactivation. Advanced functional materials for membrane modification were summarized based on their nature. Furthermore, it is suggested that membranes should be enhanced through different mechanisms mainly based on their ranks of pore size. The current review provided theoretical support regarding membrane modifications in the enhancement of virus removal and avenues for practical application.