Role of Microfiltration Membrane Morphology on Nanoparticle Purification to Enhance Downstream Purification of Viral Vectors.

In the rapidly advancing realms of gene therapy and biotechnology, the efficient purification of viral vectors is pivotal for ensuring the safety and efficacy of gene therapies. This study focuses on optimizing membrane selection for viral vector purification by evaluating key properties, including porosity, thickness, pore structure, and hydrophilicity. Notably, we employed adeno-associated virus (AAV)-sized nanoparticles (20 nm), 200 nm particles, and bovine serum albumin (BSA) to model viral vector harvesting. Experimental data from constant pressure normal flow filtration (NFF) at 1 and 2 bar using four commercial flat sheet membranes revealed distinct fouling behaviors. Symmetric membranes predominantly showed internal and external pore blockage, while asymmetric membranes formed a cake layer on the surface. Hydrophilicity exhibited a positive correlation with recovery, demonstrating an enhanced recovery with increased hydrophilicity. Membranes with higher porosity and interpore connectivity showcased superior throughput, reduced operating time, and increased recovery. Asymmetric polyether sulfone (PES) membranes emerged as the optimal choice, achieving ∼100% recovery of AAV-sized particles, an ∼44% reduction in model cell debris (200 nm particles), an ∼35% decrease in BSA, and the fastest operating time of all membranes tested. This systematic investigation into fouling behaviors and membrane properties not only informs optimal conditions for viral vector recovery but also lays the groundwork for advancing membrane-based strategies in bioprocessing.

Innovative Approaches to Poultry Processing Wastewater Treatment: The Stainless Steel Ultrafiltration Membrane as a Viable Option.

In pursuit of sustainability, we explored replacing conventional dissolved air floatation (DAF) in poultry processing wastewater (PPW) treatment with a precisely tuned 0.02 µm stainless-steel ultrafiltration (SSUF) membrane. SSUF is a robust, homogenously porous membrane with strong chemical resistance, ease of cleaning, and exceptional resistance to organic fouling. Unlike polymeric membranes, it can be regenerated multiple times, making it a cost-effective choice due to its compatibility with harsh chemical cleaning. The PPW used for the study was untreated wastewater from all processing units and post-initial screening. Our study revealed the SSUF membrane's exceptional efficiency at eliminating contaminants. It achieved an impressive removal rate of up to 99.9% for total suspended solids (TSS), oil, grease, E. coli, and coliform. Additionally, it displayed a notable reduction in chemical oxygen demand (COD), biochemical oxygen demand (BOD), and total Kjeldahl nitrogen (TKN), up to 90%, 76%, and 76%, respectively. Our investigation further emphasized the SSUF membrane's ability in pathogen removal, affirming its capacity to effectively eradicate up to 99.99% of E. coli and coliform. The measured critical flux of the membrane was 48 Lm-2h-1 at 38 kPa pressure and 1.90 m/s cross-flow velocity. In summary, our study highlights the considerable potential of the SSUF membrane. Its robust performance treating PPW offers a promising avenue for reducing its environmental impact and advocating for sustainable wastewater management practices.

Integrated Electrocoagulation, Ultrafiltration, Membrane Distillation, and Crystallization for Treating Produced Water.

Produced water (PW) generated from hydraulic fracturing operations was treated using an integrated electrocoagulation, ultrafiltration, membrane distillation, and crystallization process (EC UF MDC). The aim was to determine the viability of this integrated process for maximizing water recovery. The results obtained here indicate that optimizing the various unit operations could lead to increased recovery of PW. Membrane fouling limits all membrane separation processes. A pretreatment step to suppress fouling is essential. Here, removal of total suspended solids (TSS) and total organic carbon (TOC) was achieved by electrocoagulation (EC) followed by ultrafiltration (UF). The hydrophobic membrane used in membrane distillation may be fouled by dissolved organic compounds. Reducing membrane fouling is essential to increase the long-term durability of the membrane distillation (MD) system. In addition, combining membrane distillation with crystallization (MDC) can help reduce scale formation. By inducing crystallization in the feed tank, scale formation on the MD membrane was suppressed. The integrated EC UF MDC process can impact Water Resources/Oil & Gas Companies. Conservation of surface and groundwater is possible by treating and reusing PW. Additionally, treating PW reduces the amount of PW disposed in Class II disposal wells and promotes more environmentally sustainable operations.

Electrospun Hydrophobic Interaction Chromatography (HIC) Membranes for Protein Purification.

Responsive membranes for hydrophobic interaction chromatography have been fabricated by functionalizing poly(N-vinylcaprolactam) (PVCL) ligands on the substrate of electrospun regenerated cellulose nanofibers. Both static and dynamic binding capacities and product recovery were investigated using bovine serum albumin (BSA) and Immunoglobulin G (IgG) as model proteins. The effects of ligand chain length and chain density on static binding capacity were also studied. A static binding capacity of ~25 mg/mL of membrane volume (MV) can be achieved in optimal ligand grafting conditions. For dynamic binding studies, protein binding capacity increased with protein concentration from 0.1 to 1.0 g/L. Dynamic binding capacity increased from ~8 mg/mL MV at 0.1 g/L BSA to over 30 mg/mL at 1.0 g/L BSA. However, BSA recovery decreased as protein concentration increased from ~98% at 0.1 g/L BSA to 51% at 1 g/L BSA loading concentration. There is a clear trade-off between binding capacity and recovery rate. The electrospun substrate with thicker fibers and more open pore structures is superior to thinner fibrous membrane substrates.

High Performance Mixed-Matrix Electrospun Membranes for Ammonium Removal from Wastewaters.

Mixed-matrix electrospun membranes were developed to investigate ammonium removal from low ammonium concentration wastewaters for the first time. Particles derived from the inexpensive zeolite 13X were successfully incorporated into polyethersulfone (PES) matrices. The fabricated mixed-matrix electrospun membranes demonstrate high ammonium removal capacity reaching over 55 mg/gzeolite, more than 2.5 times higher than the previously fabricated mixed-matrix membranes via non-solvent induced phase inversion. Moreover, the membranes fabricated exhibit high permeability and ease of regeneration. Over 90% of total ammonium nitrogen (TAN) can be removed from low TAN wastewaters such as aquaculture wastewaters. In addition to zeolite 13X, other zeolite particles including zeolite Y, zeolite 3A and 4A were also incorporated into the membrane matrix. The inexpensive zeolite 13X show the highest ammonium exchange capacity. Particle type, loading and the level of its dispersion all affect TAN removal capacity.

Novel Poly(ionic liquid) Augmented Membranes for Unconventional Aqueous Phase Applications in Fractionation of Dyes and Sugar.

Poly(ionic) liquid (PIL) augmented membranes were fabricated through self-polymerization of 2-vinyl pyridine and 4-vinyl pyridine followed by dopamine triggered polymerization and bridging with inert polyamide support. The resulting membranes acquired a positive surface charge with a high degree of hydrophilicity. Fourier transformed Infra-red (FTIR) and Energy dispersive X-ray (EDX) spectroscopic investigation revealed the successful augmentation of PIL surface layer, whereas surface morphology was investigated through scanning electron microscopy (SEM) imaging. This manuscript demonstrates pi electron-induced separation of dyes with the trend in permeability: Coomassie Brilliant Blue G (CBBHG) > Remazol Brilliant Blue R (RBBR) > Eichrome Black T (EBT) > Congo Red (CR). CBBG exhibited extended conjugation over large aromatic domain. RBBR and EBT were associated withtheelectron-donating -NH2 group and electron-withdrawing -NO2 group, respectively, hence pi electron density on aromatic ring varied. The steric repulsion between two pairs of ortho hydrogens (Hs) in biphenyl moieties of CR resulted in deviation of planarity and hence aromaticity leading to the lowest permeability. The sugar fractionation followed the trend: Galactose > Mannose > Fructose > Glucose > Xylose. More hydroxyl (-OH) groups in sugars and their conformational alignment in the same direction, exhibited more lone pair of electrons leading to more interaction with PIL and hence better permeability. Pentose showed poorer permeation than hexose, whereas aldose showed better permeation than ketose.

Modeling tangential flow filtration using reverse asymmetric membranes for bioreactor harvesting.

Tangential flow filtration (TFF) has many advantages for bioreactor harvesting, as the permeate could be introduced directly to the subsequent capture step. However, membrane fouling has limited its widespread use. This is particularly problematic given the high cell densities encountered today. Here, a reverse asymmetric membrane, where the more open surface faces the feed stream and the tighter barrier layer faces the permeate stream, has been investigated. The open surface contains pores up to 40 µm in diameter while the tighter barrier layer has an average pore size of 0.4 µm. Filtration of yeast suspensions has been conducted under a range of conditions. The yeast cells are trapped in the open pore structure. The membrane stabilizes an internal porous cake that acts like a depth filter. This stabilized cake layer can remove particulate matter that would foul the barrier layer if it faced the feed stream. As filtration continues, a surface cake layer forms on the membrane surface. A resistance in series model has been developed to describe the permeate flux during TFF. The model contains three fitted parameters which can easily be determined from constant pressure normal flow filtration experiments and total recycle constant flux TFF experiments. The model can be used to estimate the capacity of the filter for a given feed stream. Our results suggest that using a reverse asymmetric membrane could avoid severe flux decline associated with fouling of the barrier layer during bioreactor harvesting.

Surface Oxidation of Ethylenechlorotrifluoroethylene (ECTFE) Membrane for the Treatment of Real Produced Water by Membrane Distillation.

Modification of ethyleneechlorotrifluoroethylene (ECTFE) membranes by simple surface oxidation was reported in the present investigation in order to induce thin hydrophilic layer on hydrophobic membrane surface for the treatment of real produced water (PW). FTIR spectra indicates the appearance of hydrophilic functional groups (⁻OH and ⁻COOH) on the membrane surface due to modification, while water contact angle, zeta potential measurement, EDX, XPS analysis confirmed the presence of O functionalized hydrophilic groups on the surface. The effect of modification temperature and the time of surface oxidation on the performance of the resulting membranes were studied systematically, which revealed that induction of optimized hydrophilicity can successfully reduce the organic fouling. However, too much hydrophilic surface induces polar/electrostatic interaction resulting salt deposition on membrane surface. A simple on site cleaning procedure was demonstrated to be successful for the treatment PW for at least three consecutive cycles of membrane distillation (MD).

Identification and Characterization of Novel Fc-Binding Heptapeptides from Experiments and Simulations.

Purification of biologically-derived therapeutics is a major cost contributor to the production of this rapidly growing class of pharmaceuticals. Monoclonal antibodies comprise a large percentage of these products, therefore new antibody purification tools are needed. Small peptides, as opposed to traditional antibody affinity ligands such as Protein A, may have advantages in stability and production costs. Multiple heptapeptides that demonstrate Fc binding behavior that have been identified from a combinatorial peptide library using M13 phage display are presented herein. Seven unique peptide sequences of diverse hydrophobicity and charge were identified. All seven peptides showed strong binding to the four major human IgG isotypes, human IgM, as well as binding to canine, rat, and mouse IgG. These seven peptides were also shown to bind human IgG4 from DMEM cell culture media with 5% FCS and 5 g/L ovalbumin present. These peptides may be useful as surface ligands for antibody detection and purification purposes. Molecular docking and classical molecular dynamics (MD) simulations were conducted to elucidate the mechanisms and energetics for the binding of these peptides to the Fc region. The binding site was found to be located between the two glycan chains inside the Fc fragment. Both hydrogen bonding and hydrophobic interactions were found to be crucial for the binding interactions. Excellent agreement for the binding strength was obtained between experimental results and simulations.

Specificity in cationic interaction with poly(N-isopropylacrylamide).

Classical molecular dynamics (MD) simulations were conducted for PNIPAM in 1 M monovalent alkali chloride salt solutions as well as in 0.5 M divalent Mg(2+) and Ca(2+) chloride salt solutions. It was found that the strength for the direct alkali ion-amide O binding is strongly correlated with the size of the ionic radius. The smallest Li(+) ion binds strongest to amide O, and the largest Cs(+) ion has the weakest interaction with the amide bond. For the divalent Mg(2+) and Ca(2+) ions, their interactions with the amide bond are weak and appear to be mediated by the water molecules, particularly in the case of Mg(2+), resulting from their strong hydration. The direct binding between the cations and amide O requires partial desovlation of the ions that is energetically unfavorable for Mg(2+) and also to a great extent for Ca(2+). The higher cation charge makes the electrostatic interaction more favorable but the dehydration process less favorable. This competition between electrostatic interaction and the dehydration process largely dictates whether the direct binding between the cation and amide O is energetically preferred or not. For monovalent alkali ions, it is energetically preferred to bind directly with the amide O. Moreover, Li(+) ion is also found to associate strongly with the hydrophobic residues on PNIPAM.

The effects of chemical substitution and polymerization on the pKa values of sulfonic acids.

The effects of ring substitution on the pK(a) value of benzenesulfonic acid (BSA) were investigated using a combined quantum mechanical and classical approach. Ring substitution with strong electron-withdrawing elements such as F, Cl, and Br is found to enhance the acidity of the BSA. More importantly, ring substitution with -NO(2) groups which form an extended conjugated pi-system with the benzene ring exhibits the strongest enhancement of the acidity. The effects of polymerization on the styrenesulfonic acid (SSA) were also investigated by solving the classical Poisson-Boltzmann equation. It is found that polymerization significantly decreases the acidity of SSA due to the alteration of the electrostatic environment surrounding the acid group upon polymerization. The average pK(a) value converges to 2.9 from the corresponding monomer value of -0.53 at a degree of polymerization of 8-12. These results shed significant light on how to design sulfonic-acid-based solid acid catalysts to achieve desired catalytic properties.