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Membranes
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Bioprocessing with Membranes: Filtration and Chromatography
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Bioprocessing with Membranes: Filtration and Chromatography

A special issue of Membranes (ISSN 2077-0375). This special issue belongs to the section "Membrane Processing and Engineering".

Deadline for manuscript submissions: closed (15 March 2023) | Viewed by 27700

Special Issue Editor

Dr. Marília Mateus

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Guest Editor
iBB—Institute for Bioengineering and Biosciences, IST, Universidade de Lisboa, Av. Rovisco Pais, nº1, 1049-001 Lisbon, Portugal
Interests: bioprocess engineering; biopharmaceuticals; bioseparations; bioreactors; membrane processes
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

Membrane filtration has been widely considered a useful separation technique, mainly due to the advantages it presents when compared to other separation techniques, both economically and regarding the quality of the recovered products. Applications can be found in a wide variety of industries, such as wastewater treatment or water production/reclamation, biorefineries, food and beverage, and biopharmaceutical sectors. The main purposes are removal of dispersions, pollutants or impurities, concentration of target products and recovery of byproducts from process streams, and retention of cells or other biocatalysts in continuous or perfused bioreactors. Major concerns in membrane filtration processes are still the impact of membrane fouling on the process performance, the implications of not selecting the best operation conditions or sequence of processes, and the energy and membrane cost requirements for processing. Which innovative strategies are thought to overcome these difficulties?

Chromatography is a fine-tuning separation method important for many bioprocesses, in which specific physical–chemical and affinity properties of the target products and of the respective impurities are addressed to meet effective separation and high purity in a final product when required. Membranes have been used as chromatographic solid matrices (e.g., adsorbers), particularly for the purification and polishing steps of large biomolecules, such as biopharmaceuticals. Which new membrane adsorbers are being developed or need development? Which novelties are proposed for membrane module design? Modeling and applications of continuous chromatography operation and the outputs of membrane column arrangements would probably be very insightful.

This Special Issue aims to cover the latest developments and innovations regarding membrane filtration and membrane chromatography operation and their systems for bioprocessing. Potential topics include but are not limited to the following:

  • Fundamentals and applications—new approaches
  • Batch and continuous processing strategies
  • Up- and downstream process integration
  • Membrane improvement for fouling control
  • Development of membrane adsorbers
  • Depth filtration with membranes
  • Comparative performance of membrane materials/structures
  • Membrane bioreactors
  • Scale-up studies and scale-down devices
  • Module design
  • Process economics
  • Modeling of membrane performance and membrane systems

Dr. Marília Mateus
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Membranes is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Published Papers (11 papers)

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Research

16 pages, 9582 KiB  
Article
Scaling Up of Steric Exclusion Membrane Chromatography for Lentiviral Vector Purification
by Jennifer Julia Labisch, Richard Paul, G. Philip Wiese and Karl Pflanz
Membranes 2023, 13(2), 149; https://doi.org/10.3390/membranes13020149 - 24 Jan 2023
Cited by 3 | Viewed by 3398
Abstract
Lentiviral vectors (LVs) are widely used in clinical trials of gene and cell therapy. Low LV stability incentivizes constant development and the improvement of gentle process steps. Steric exclusion chromatography (SXC) has gained interest in the field of virus purification but scaling up [...] Read more.
Lentiviral vectors (LVs) are widely used in clinical trials of gene and cell therapy. Low LV stability incentivizes constant development and the improvement of gentle process steps. Steric exclusion chromatography (SXC) has gained interest in the field of virus purification but scaling up has not yet been addressed. In this study, the scaling up of lentiviral vector purification by SXC with membrane modules was approached. Visualization of the LVs captured on the membrane during SXC showed predominant usage of the upper membrane layer. Furthermore, testing of different housing geometries showed a strong influence on the uniform usage of the membrane. The main use of the first membrane layer places a completely new requirement on the scaling of the process and the membrane modules. When transferring the SXC process to smaller or larger membrane modules, it became apparent that scaling of the flow rate is a critical factor that must be related to the membrane area of the first layer. Performing SXC at different scales demonstrated that a certain critical minimum surface area-dependent flow rate is necessary to achieve reproducible LV recoveries. With the presented scaling approach, we were able to purify 980 mL LVs with a recovery of 68%. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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22 pages, 3359 KiB  
Article
Adsorption Performance of a Multimodal Anion-Exchange Chromatography Membrane: Effect of Liquid Phase Composition and Separation Mode
by Tomáš Kurák and Milan Polakovič
Membranes 2022, 12(12), 1173; https://doi.org/10.3390/membranes12121173 - 22 Nov 2022
Cited by 1 | Viewed by 1897
Abstract
Membrane chromatography is a modern, high-throughput separation method that finds important applications in therapeutic protein purification. Multimodal, salt-tolerant membranes are the most recent innovation in chromatographic membrane adsorbents. Due to the complex structure of their ligands and the bimodal texture of their carriers, [...] Read more.
Membrane chromatography is a modern, high-throughput separation method that finds important applications in therapeutic protein purification. Multimodal, salt-tolerant membranes are the most recent innovation in chromatographic membrane adsorbents. Due to the complex structure of their ligands and the bimodal texture of their carriers, their adsorption properties have not been sufficiently investigated. This work deals with the equilibrium and kinetic properties of a multimodal anion-exchange chromatography membrane, Sartobind STIC. Single- and two-component adsorption experiments were carried out with bovine serum albumin (BSA) and salmon DNA as model target and impurity components. The effect of the Hofmeister series ions and ionic strength on the BSA/DNA adsorption was investigated in micromembrane flow experiments. A significant difference was observed between the effects of monovalent and polyvalent ions when strong kosmotropic salts with polyvalent anions acted as strong displacers of BSA. On the contrary, DNA binding was rather high at elevated ionic strength, independent of the salt type. Two-component micromembrane experiments confirmed very high selectivity of DNA binding at a rather low sodium sulfate feed content and at pH 8. The strength of binding was examined in more than a dozen different desorption experiments. While BSA was desorbed relatively easily using high salt concentrations independent of buffer type and pH, while DNA was desorbed only in a very limited measure under any conditions. Separation experiments in a laboratory membrane module were carried out for the feed containing 1 g/L of BSA, 0.3 g/L of DNA, and 0.15 M of sodium sulfate. The negative flow-through mode was found to be more advantageous than the bind-elute mode, as BSA was obtained with 99% purity and a 97% yield. Membrane reuse was investigated in three adsorption-desorption-regeneration cycles. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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16 pages, 3841 KiB  
Article
Scalable, Robust and Highly Productive Novel Convecdiff Membrane Platform for mAb Capture
by Mario Grünberg, Kim B. Kuchemüller, Katrin Töppner and Ricarda A. Busse
Membranes 2022, 12(7), 677; https://doi.org/10.3390/membranes12070677 - 30 Jun 2022
Cited by 6 | Viewed by 2232
Abstract
The recombinant monoclonal antibody capture step represents the current bottleneck in downstream processing. Protein A resins are diffusion-limited chromatography materials which require low flow rates to achieve a binding capacity above 30 g L−1 with the result of low productivity. Here, we [...] Read more.
The recombinant monoclonal antibody capture step represents the current bottleneck in downstream processing. Protein A resins are diffusion-limited chromatography materials which require low flow rates to achieve a binding capacity above 30 g L−1 with the result of low productivity. Here, we present a novel chromatography membrane combining superior binding capacities with high flow rates for high productivity while achieving comparable product quality as state-of-the-art protein A resins. Further, we demonstrate full scalability of this convecdiff technology with experimental data demonstrating suitability for bioprocessing at different scales. This technology results in more than 10-fold higher productivity compared to Protein A resins, which is maintained during scale up. We demonstrate the influence of residence times, feed titers and the cleaning regime on productivity and indicate optimal utilization of the convecdiff membrane based on feed titer availability. The underlying high productivity and short cycle times of this material enable the purification of monoclonal antibodies with 10-times less chromatography material used per batch and utilization of the membrane within one batch. Provided in disposable consumables, this novel technology will remove column handling in bioprocesses and resin re-use over multiple batches. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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10 pages, 2126 KiB  
Article
Comparative Evaluation of the Performance of Sterile Filters for Bioburden Protection and Final Fill in Biopharmaceutical Processes
by Jimin Na, Dongwoo Suh, Young Hoon Cho and Youngbin Baek
Membranes 2022, 12(5), 524; https://doi.org/10.3390/membranes12050524 - 16 May 2022
Cited by 3 | Viewed by 2785
Abstract
Sterile filtration processes are widely used in the production of biotherapeutics for microorganism removal and product sterility. Sterile filtration processes can be applied to buffer preparation and cell culture media preparation in biotherapeutics processes, and to final sterilization or final filling in downstream [...] Read more.
Sterile filtration processes are widely used in the production of biotherapeutics for microorganism removal and product sterility. Sterile filtration processes can be applied to buffer preparation and cell culture media preparation in biotherapeutics processes, and to final sterilization or final filling in downstream processes. Owing to their broad range of applications in bioprocessing, various 0.2/0.22 μm sterile filters with different polymer materials (i.e., hydrophilic PVDF and PES) and nominal pore sizes are commercially available. The objective of this study was to evaluate two different commercial sterile filters in terms of filtration performance in various sterile filtration processes of biopharmaceutical production. The results demonstrate the importance of choosing the appropriate filter considering the process type and target removal/transport product to ensure efficient sterile filtration in the production of biotherapeutics. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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12 pages, 4638 KiB  
Communication
The Suitability of Latex Particles to Evaluate Critical Process Parameters in Steric Exclusion Chromatography
by Friederike Eilts, Marleen Steger, Keven Lothert and Michael W. Wolff
Membranes 2022, 12(5), 488; https://doi.org/10.3390/membranes12050488 - 30 Apr 2022
Cited by 6 | Viewed by 1860
Abstract
The steric exclusion chromatography (SXC) is a rather new method for the purification of large biomolecules and biological nanoparticles based on the principles of precipitation. The mutual steric exclusion of a nonionic organic polymer, i.e., polyethylene glycol (PEG), induces target precipitation and leads [...] Read more.
The steric exclusion chromatography (SXC) is a rather new method for the purification of large biomolecules and biological nanoparticles based on the principles of precipitation. The mutual steric exclusion of a nonionic organic polymer, i.e., polyethylene glycol (PEG), induces target precipitation and leads to their retention on the chromatographic stationary phase. In this work, we investigated the application of latex particles in the SXC by altering the particle’s surface charge as well as the PEG concentration and correlated both with their aggregation behavior. The parameters of interest were offline precipitation kinetics, the product recovery and yield, and the chromatographic column blockage. Sulfated and hydroxylated polystyrene particles were first characterized concerning their aggregation behavior and charge in the presence of PEG and different pH conditions. Subsequently, the SXC performance was evaluated based on the preliminary tests. The studies showed (1) that the SXC process with latex particles was limited by aggregation and pore blockage, while (2) not the aggregate size itself, but rather the aggregation kinetics dominated the recoveries, and (3) functionalized polystyrene particles were only suitable to a limited extent to represent biological nanoparticles of comparable size and charge. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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17 pages, 2764 KiB  
Article
Evaluation of Host Cell Impurity Effects on the Performance of Sterile Filtration Processes for Therapeutic Viruses
by Evan Wright, Karina Kawka, Maria Fe C. Medina and David R. Latulippe
Membranes 2022, 12(4), 359; https://doi.org/10.3390/membranes12040359 - 24 Mar 2022
Cited by 3 | Viewed by 2733
Abstract
Efficient downstream processing represents a significant challenge in the rapidly developing field of therapeutic viruses. While it is known that the terminal sterile filtration step can be a major cause of product loss, there is little known about the effect of host cell [...] Read more.
Efficient downstream processing represents a significant challenge in the rapidly developing field of therapeutic viruses. While it is known that the terminal sterile filtration step can be a major cause of product loss, there is little known about the effect of host cell impurities (DNA and protein) on filtration performance. In this study, fractions of relatively pure Vero host cell protein and DNA were spiked into a highly pure preparation of vesicular stomatitis virus (VSV). Then, the resulting solutions were sterile filtered using two commercially available 0.22 µm rated microfiltration membranes. A combination of transmembrane pressure measurements, virus recovery measurements, and post-filtration microscopy images of the microfiltration membranes was used to evaluate the sterile filtration performance. It was found that increasing the amount of host cell protein from approximately 1 µg/mL (in the un-spiked VSV preparation) to 25 µg/mL resulted in a greater extent of membrane fouling, causing the VSV recovery to decrease from 89% to 65% in experiments conducted with the highly asymmetric Express PLUS PES membrane and to go as low as 48% in experiments conducted with the symmetric Durapore PVDF membrane. Similar effects were not seen when bovine serum albumin, a common model protein used in filtration studies, was spiked into the VSV preparation, which indicates that the sterile filtration performance is critically dependent on the complex composition of the mixture of host cell proteins rather than the presence of any protein. The results presented in this work provide important insights into the role of host cell impurities on the performance of sterile filtration processes for therapeutic viruses. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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14 pages, 1999 KiB  
Article
Membrane Bioreactor for Simultaneous Synthesis and Fractionation of Oligosaccharides
by Vanessa A. Botelho, Marília Mateus, José C. C. Petrus and Maria Norberta de Pinho
Membranes 2022, 12(2), 171; https://doi.org/10.3390/membranes12020171 - 31 Jan 2022
Cited by 7 | Viewed by 2214
Abstract
Galacto-oligosaccharides (GOS) are prebiotic sugars obtained enzymatically from lactose and used in food industry due to their nutritional advantages or technological properties. Selective mass transport and enzymatic synthesis were integrated and followed using a membrane bioreactor, so that selective removal of reaction products [...] Read more.
Galacto-oligosaccharides (GOS) are prebiotic sugars obtained enzymatically from lactose and used in food industry due to their nutritional advantages or technological properties. Selective mass transport and enzymatic synthesis were integrated and followed using a membrane bioreactor, so that selective removal of reaction products may lead to increased conversions of product-inhibited or thermodynamically unfavorable reactions. GOS syntheses were conducted on lactose solutions (150 g·L−1) at 40 °C and 10 Uβ-galactosidase.mL−1, and sugar fractionation was performed by cellulose acetate membranes. Effects of pressure (20; 24 bar) and crossflow velocity (1.7; 2.0; 2.4 m·s−1) on bioreactor performance were studied. Simultaneous GOS synthesis and production fractionation increased GOS production by 60%, in comparison to the same reactions promoted without permeation. The presence of a high-molecular-weight solute, the enzyme, in association with high total sugar concentration, leads to complex selective mass transfer characteristics. Without the enzyme, the membrane presented tight ultrafiltration characteristics, permeating mono- and disaccharides and retaining just 25% of trisaccharides. During simultaneous synthesis and fractionation, GOS-3 were totally retained, and GOS-2 and monosaccharides were retained at 80% and 40%, respectively. GOS synthesis—hydrolysis evolution was strongly dependent on crossflow velocity at 20 bar but became fairly independent at 24 bar. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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13 pages, 4962 KiB  
Article
Liposome Sterile Filtration Characterization via X-ray Computed Tomography and Confocal Microscopy
by Thomas F. Johnson, Kyle Jones, Francesco Iacoviello, Stephen Turner, Nigel B. Jackson, Kalliopi Zourna, John H. Welsh, Paul R. Shearing, Mike Hoare and Daniel G. Bracewell
Membranes 2021, 11(11), 905; https://doi.org/10.3390/membranes11110905 - 22 Nov 2021
Cited by 1 | Viewed by 2318
Abstract
Two high resolution, 3D imaging techniques were applied to visualize and characterize sterilizing grade dual-layer filtration of liposomes, enabling membrane structure to be related with function and performance. Two polyethersulfone membranes with nominal retention ratings of 650 nm and 200 nm were used [...] Read more.
Two high resolution, 3D imaging techniques were applied to visualize and characterize sterilizing grade dual-layer filtration of liposomes, enabling membrane structure to be related with function and performance. Two polyethersulfone membranes with nominal retention ratings of 650 nm and 200 nm were used to filter liposomes of an average diameter of 143 nm and a polydispersity index of 0.1. Operating conditions including differential pressure were evaluated. X-ray computed tomography at a pixel size of 63 nm was capable of resolving the internal geometry of each membrane. The respective asymmetry and symmetry of the upstream and downstream membranes could be measured, with pore network modeling used to identify pore sizes as a function of distance through the imaged volume. Reconstructed 3D digital datasets were the basis of tortuous flow simulation through each porous structure. Confocal microscopy visualized liposome retention within each membrane using fluorescent dyes, with bacterial challenges also performed. It was found that increasing pressure drop from 0.07 MPa to 0.21 MPa resulted in differing fluorescent retention profiles in the upstream membrane. These results highlighted the capability for complementary imaging approaches to deepen understanding of liposome sterilizing grade filtration. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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12 pages, 3006 KiB  
Article
The Pilot Study of the Influence of Free Ammonia on Membrane Fouling during the Partial Nitrosation of Pig Farm Anaerobic Digestion Liquid
by Hanxiao Bian, Zhiping Zhu, Qianwen Sui and Shunli Wang
Membranes 2021, 11(11), 894; https://doi.org/10.3390/membranes11110894 - 19 Nov 2021
Viewed by 1602
Abstract
The problem of membrane fouling is a key factor restricting the application of the membrane bioreactor (MBR) in the partial nitrosation (PN) and anaerobic ammonia oxidation (anammox) processes. In this study, the pilot-scale continuous flow MBR was used to start up the partial [...] Read more.
The problem of membrane fouling is a key factor restricting the application of the membrane bioreactor (MBR) in the partial nitrosation (PN) and anaerobic ammonia oxidation (anammox) processes. In this study, the pilot-scale continuous flow MBR was used to start up the partial nitrosation process in order to investigate the change trend of mid-transmembrane pressure (TMP) in the process of start-up, which was further explored to clarify the membrane fouling mechanism in the pilot-scale reactor. The results showed that the MBR system was in a stable operating condition during the partial nitrosation operation and that the online automatic backwash operation mode is beneficial in alleviating membrane fouling and reducing the cost of membrane washing. Particular attention was paid to the influence trend of free ammonia (FA)on membrane fouling, and it was found that the increase in FA concentration plays the most critical role in membrane fouling. The increase in FA concentration led to an increase in the extracellular polymer (EPS), dissolved microorganism product (SMP) and soluble chemical oxygen demand (SCOD) concentration. FA was extremely significantly correlated with EPS and SCOD, and the FA concentration was approximately 20.7 mg/L. The SCODeff (effluent SCOD concentration) concentration was approximately 147 mg/L higher than the SCODinf (influent SCOD concentration) concentration. FA mainly affects membrane fouling by affecting the concentration of EPS and SCOD. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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20 pages, 2811 KiB  
Article
The Use of NaOH Solutions for Fouling Control in a Membrane Bioreactor: A Feasibility Study
by Wirginia Tomczak, Ireneusz Grubecki and Marek Gryta
Membranes 2021, 11(11), 887; https://doi.org/10.3390/membranes11110887 - 18 Nov 2021
Cited by 9 | Viewed by 2348
Abstract
Nowadays, the microbial production of 1,3-propanediol (1,3-PD) is recognized as preferable to the chemical synthesis. However, finding a technological approach allowing the production of 1,3-PD in the membrane bioreactor (MBR) is a great challenge. In the present study, a ceramic ultrafiltration (UF) membrane [...] Read more.
Nowadays, the microbial production of 1,3-propanediol (1,3-PD) is recognized as preferable to the chemical synthesis. However, finding a technological approach allowing the production of 1,3-PD in the membrane bioreactor (MBR) is a great challenge. In the present study, a ceramic ultrafiltration (UF) membrane (8 kDa) for treatment of 1,3-PD broths was used. It has been demonstrated that the membrane used provides the stable permeate flux that is necessary to ensure the stability of the fermentation process in MBR technology. It was noticed that the broth pH has a significant impact on both the final 1,3-PD concentration and permeate flux. Moreover, the feasibility of using NaOH for fouling control in the MBR was evaluated. It has been shown that 1% NaOH solution is effective in restoring the initial membrane performance. To the best of our knowledge, this study is the first to shed light onto the possibility of reducing the amount of the alkaline solutions generated during the MBR operation. Indeed, it has been found that 1% NaOH solution can be successfully used several times for both membrane cleaning and to stabilize the broth pH. Finally, based on the results obtained, the technological conceptions of the MBR technology were designed. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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11 pages, 3157 KiB  
Article
Effects of Impurities from Sugar Excipient on Filtrate Flux during Ultrafiltration and Diafiltration Process
by Jieun Lee, Jiwon Na and Youngbin Baek
Membranes 2021, 11(10), 775; https://doi.org/10.3390/membranes11100775 - 11 Oct 2021
Cited by 2 | Viewed by 2582
Abstract
Sugar excipients such as sucrose and maltose are widely used for biopharmaceutical formulation to improve protein stability and to ensure isotonicity for administration. However, according to recent literature, pharmaceutical-grade sucrose contained nanoparticulate impurities (NPIs) that result in protein aggregation and degradation. The objective [...] Read more.
Sugar excipients such as sucrose and maltose are widely used for biopharmaceutical formulation to improve protein stability and to ensure isotonicity for administration. However, according to recent literature, pharmaceutical-grade sucrose contained nanoparticulate impurities (NPIs) that result in protein aggregation and degradation. The objective of this study was to evaluate the filtrate flux behavior of sugar solution during ultrafiltration (UF) and diafiltration (DF). Filtrate flux data were obtained using either a tangential flow filtration (TFF) system for DF experiments or a normal flow filtration system for UF experiments. In diafiltration experiments, which were performed using 7 g/L of human immunoglobulin G in a 20 mM histidine buffer with the 100 mM sucrose or maltose, the filtrate flux with sucrose solution decreased significantly. In contrast, the one with maltose solution was in good correspondence with the calculated filtrate flux accounting for the effects of solution viscosity. This large decline in the flux was also observed during UF experiments, in which the presence of NPIs was identified by dynamic light scattering analysis and by capturing an SEM image of the membrane surface after filtration. In addition, highly purified sucrose resulted in a much lower flux decline in TFF in the absence of NPIs. These results provide important insights into the factors governing the optimization of the UF/DF process using appropriate excipients for biopharmaceutical formulation. Full article
(This article belongs to the Special Issue Bioprocessing with Membranes: Filtration and Chromatography)
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