Remove legacy perfluoroalkyl acids and emerging per- and polyfluoroalkyl ether acids by single-use and regenerable anion exchange resins: Rapid small-scale column tests and model fits.

Rapid small-scale column tests (RSSCT) are used to study the removal of per- and polyfluoroalkyl substances (PFAS) for drinking water treatment by ion exchange. Breakthroughs of 15 emerging per- and perfluoroalkyl ether acids and six legacy perfluoroalkyl acid analogs are studied using a single-use PFAS-selective anion exchange resin (AER1) and a regenerable, generic anion exchange resin (AER2). The Bohart-Adams model was used to describe and predict breakthrough, with the modeled results reasonably aligned with RSSCT results in most cases, enabling shorter RSSCT duration for future applications. AER1 exhibited high uptake capacity with no breakthrough for 11 of the 21 tested PFAS during the 144,175 BV continuous operation, allowing compliance with the new National Primary Drinking Water Regulation in many application scenarios. AER2 exhibited much faster breakthroughs for most PFAS and is not a promising option for drinking water treatment. However, the summed PFAS capacity via model fit and total PFAS adsorbed via measurement were only <0.01 % of both resin capacities at full breakthrough, suggesting PFAS could only occupy a tiny portion of the ion exchange sites even for the PFAS-selective AER1. Ether group insertion in the PFAS group leads to later breakthrough, and linear isomers were better captured by the resins than the branched isomers. Overall, PFAS uptake capacity increases and kinetics decrease when the PFAS molecular volume increases. Regeneration using 10 % NaCl solutions partially released PFAS from AER2 but not from AER1, with more short-chain PFAS released than long-chain ones. Ether group insertion decreased the PFAS recoveries during the regeneration of AER2. The regenerated resins showed much faster breakthroughs than the pristine resins, making them unfavorable for drinking water treatment applications. Adsorption displacement of short-chain PFAS by long-chain PFAS was observed in pristine AER1, and post-regeneration leaching occurred for both resins, both phenomena making the resins a possible PFAS source in long-term use.

Removal of copper and iron from ethanolic solutions by an anion exchange resin and its implication to rare-earth magnet recycling.

In the recycling of end-of-life rare-earth magnets, the recovery of non-rare earth constituents is often neglected. In the present study, strong cation and anion exchange resins were tested batchwise for the recovery of the non-rare-earth constituents of permanent magnets (copper, cobalt, manganese, nickel and iron) from synthetic aqueous and ethanolic solutions. The cation exchange resin recovered most of metal ions from aqueous and ethanolic feeds, whereas the anion exchange resin could selectively recover copper and iron from ethanolic feeds. The highest uptake of iron and copper was found for 80 vol% and 95 vol% multi-element ethanolic feeds, respectively. A similar trend in selectivity of the anion resin was observed in breakthrough curve studies. Batch experiments, UV-Vis, FT-IR and XPS studies were performed to elucidate the ion exchange mechanism. The studies indicate that the formation of chloro complexes of copper and their exchange by the (hydrogen) sulfate counter ions of the resin have an important role in the selective uptake of copper from the 95 vol% ethanolic feed. Iron(II) was largely oxidized to iron(III) in ethanolic solutions and was expected to be recovered by the resin in the form of iron(II) and iron(III) complexes. The moisture content of the resin did not have a significant role on the selectivity for copper and iron.

Adsorptive Removal of Direct Azo Dyes from Textile Wastewaters Using Weakly Basic Anion Exchange Resin.

Direct dyes are still widely used for coloring a variety of materials due to their ease of use and the wide range of colors available at a moderate cost of production. In the aquatic environment, some direct dyes, especially the azo type and their biotransformation products, are toxic, carcinogenic and mutagenic. Hence the need for their careful removal from industrial effluents. It was proposed adsorptive retention of C.I. Direct Red 23 (DR23), C.I. Direct Orange 26 (DO26) and C.I. Direct Black 22 (DB22) from effluents using anion exchange resin of tertiary amine functionalities Amberlyst A21 (A21). Applying the Langmuir isotherm model, the monolayer capacities were calculated as 285.6 mg/g for DO26 and 271.1 mg/g for DO23. The Freundlich isotherm model seems to be the better one for the description of DB22 uptake by A21, and the isotherm constant was found to be 0.609 mg1-1/n L1/n/g. The kinetic parameters revealed that the pseudo-second-order model could be used for the description of experimental data rather than the pseudo-first-order model or intraparticle diffusion model. The dye adsorption decreased in the presence of anionic and non-ionic surfactants, while their uptake was enhanced in the presence of Na2SO4 and Na2CO3. Regeneration of the A21 resin was difficult; a slight increase in its efficiency was observed using 1M HCl, 1 M NaOH and 1 M NaCl solutions in 50% v/v methanol.

Removal and destruction of perfluoroalkyl ether carboxylic acids (PFECAs) in an anion exchange resin and electrochemical oxidation treatment train.

Perfluoroalkyl ether carboxylic acids (PFECAs) are a group of emerging recalcitrant contaminants that are being developed to replace legacy per- and polyfluoroalkyl substances (PFAS) in industrial applications and that are generated as by-products in fluoropolymer manufacturing. Here, we report on the removal and destruction of four structurally different PFECAs using an integrated anion exchange resin (AER) and electrochemical oxidation (ECO) treatment train. Results from this work illustrated that (1) flow-through columns packed with PFAS-selective AERs are highly effective for the removal of PFECAs and (2) PFECA affinity is strongly correlated with their hydrophobic features. Regeneration of the spent resin columns revealed that high percentage (e.g., 80%) of organic cosolvent is necessary for achieving 60-100% PFECA release, and regeneration efficiency was higher for a macroporous resin than a gel-type resin. Treatment of spent regenerants showed (1) >99.99% methanol removal was achieved by distillation, (2) >99.999% conversion of the four studied PFECAs was achieved during the ECO treatment of the still bottoms after 24 hours with an energy per order of magnitude of PFECA removal (EE/O) <1.03 kWh/m3 of total groundwater treated, and (3) >85% of the organic fluorine was recovered as inorganic fluoride. Trifluoroacetic acid (TFA), perfluoropropionic acid (PFPrA), and perfluoro-2-methoxyacetic acid (PFMOAA) were confirmed via high-resolution mass spectrometry as transformation products (TPs) in the treated still bottoms, and two distinctive degradation schemes and four reaction pathways are proposed for the four PFECAs. Lastly, dissolved organic matter (DOM) inhibited uptake, regeneration, and oxidation of PFECAs throughout the treatment train, suggesting pretreatment steps targeting DOM removal can enhance the system's treatment efficiency. Results from this work provide guidelines for developing effective separation-concentration-destruction treatment trains and meaningful insights for achieving PFECA destruction in impacted aquatic systems.

Degradation of the mixed nuclear-grade cationic and anionic exchange resins using Fe2+/H+ homogeneous Fenton oxidation.

To further improve the treatment capacity of actual wastes, H+ was introduced into the homogeneous Fenton system as a co-catalyst for dissolution and degradation of the mixed nuclear-grade cationic and anionic exchange resins. The effects of acid type and concentration, catalyst type and concentration, H2O2 dosage, initial temperature, antifoaming agent and resin ratio were studied. The concentration of inorganic acid, type and concentration of catalyst had significant influence on the decomposition of mixed resins. The experimental results showed that when the mixing ratio of resins was 1:1, the initial temperature was 96 ± 1 °C, the amount of H2O2 was 200 mL, and the concentration of H+/Fe2+ was 1 M/0.1 M, complete dissolution and 79% weight reduction of mixed resins were obtained. Combined with density functional theory (DFT) calculations, cationic exchange resin and anionic exchange resin showed different reactivity in the experiment. Hydroxyl radicals (•OH) tended to attack -SO3- groups with more negative charges, and the barrier energy of -SO3- ion dissociation was 8.2 kcal/mol, which caused the cationic exchange resin to be easily destroyed. According to the characterization results, the characteristic intermediates were determined, indicating that desulfonation, valence change of nitrogen atom, and cleavage of long-chain carbon skeleton existed during the reaction, but incomplete oxidation still remained.

Gas-Free Continuously Regenerated Impurity Removal Device for Ion Chromatography.

A gas-free continuously regenerated-anion impurity removal device (CR-ARD) is described for ion chromatography (IC). It is a sandwiched configuration consisting of three channels. The central eluent channel is isolated from two outer regenerant channels by stacked anion exchange membranes (sAEM) and a bipolar membrane (BPM) plus stacked cation exchange membranes (BPM-sCEM), in which the anion exchange side of BPM is facing the central channel. The sAEM side is an anode with respect to the cathode of the BPM-sCEM side. The central channel is packed with strongly basic anion exchange resins in hydroxide form. The application of an electrical voltage to the device causes enhanced water splitting at the interface of BPM to occur and the potential drives the hydroxide ions through the resin phase toward the anode, thereby enabling continuous regeneration of the resin and avoiding any off-line regeneration. Meanwhile, nonhydroxide anionic impurities (basically carbonate) are transported toward the anode through the sAEM. It does not produce gas in the eluent channel, and no degasser is required afterward. It shows a strong ability to remove anionic impurities, as indicated by effective removal of 5 mM CO32- injected sample plug (25 µL, 125 pmol) or continuous removal of 600 µM KNO3 solution at 1 mL/min (10 nmol/s) over 300 min. Much improved peak integration at the gradient mode and a higher signal-to-noise ratio at the isocratic mode can be achieved by CR-ARD. To our knowledge, this is the first account of gas-free CR-ARD.

Precision analysis for the determination of steric mass action parameters using eight tobacco host cell proteins.

Plants are advantageous as biopharmaceutical manufacturing platforms because they allow the economical and scalable upstream production of proteins, including those requiring post-translational modifications, but do not support the replication of human viruses. However, downstream processing can be more labor-intensive compared to fermenter-based systems because the product is often mixed with abundant host cell proteins (HCPs). Modeling chromatographic separation can minimize the number of process development experiments and thus reduce costs. An important part of such modeling is the sorption isotherm, such as the steric mass action (SMA) model, which describes the multicomponent protein-salt equilibria established in ion-exchange systems. Here we purified ten HCPs, including 2-Cys-peroxiredoxin, from tobacco (Nicotiana tabacum and N. benthamiana). For eight of these HCPs, we obtained sufficient quantities to determine the SMA binding parameters (KSMA and ν) under different production-relevant conditions. We studied the parameters for 2-Cys-peroxiredoxin on Q-Sepharose HP in detail, revealing that pH, resin batch and buffer batch had little influence on KSMA and ν, with coefficients of variation (COVs) less than 0.05 and 0.21, respectively. In contrast, the anion-exchange resins SuperQ-650S, Q-Sepharose FF and QAE-550C led to COVs of 0.69 for KSMA and 0.05 for ν, despite using the same quaternary amine functional group as Q-Sepharose HP. Plant cultivation in summer vs winter resulted in COVs of 0.09 for KSMA and 0.02 for ν, revealing a small impact compared to COVs of 17.15 for KSMA and 0.20 for ν when plants were grown in different settings (climate-controlled phytotron vs greenhouse). We conclude that plant cultivation can substantially affect protein properties and the resulting SMA parameters. Accordingly, plant growth but also protein purification and characterization for chromatography model building should be tightly controlled and well documented.

Analyses of Inositol Phosphates and Phosphoinositides by Strong Anion Exchange (SAX)-HPLC.

The phosphate esters of myo-inositol (Ins) occur ubiquitously in biology. These molecules exist as soluble or membrane-resident derivatives and regulate a plethora of cellular functions including phosphate homeostasis, DNA repair, vesicle trafficking, metabolism, cell polarity, tip-directed growth, and membrane morphogenesis. Phosphorylation of all inositol hydroxyl groups generates phytic acid (InsP6), the most abundant inositol phosphate present in eukaryotic cells. However, phytic acid is not the most highly phosphorylated naturally occurring inositol phosphate. Specialized small molecule kinases catalyze the formation of the so-called myo-inositol pyrophosphates (PP-InsPs), such as InsP7 and InsP8. These molecules are characterized by one or several "high-energy" diphosphate moieties and are ubiquitous in eukaryotic cells. In plants, PP-InsPs play critical roles in immune responses and nutrient sensing. The detection of inositol derivatives in plants is challenging. This is particularly the case for inositol pyrophosphates because diphospho bonds are labile in plant cell extracts due to high amounts of acid phosphatase activity. We present two steady-state inositol labeling-based techniques coupled with strong anion exchange (SAX)-HPLC analyses that allow robust detection and quantification of soluble and membrane-resident inositol polyphosphates in plant extracts. These techniques will be instrumental to uncover the cellular and physiological processes controlled by these intriguing regulatory molecules in plants.

Facile and Safe Synthesis of Novel Self-Pored Amine-Functionalized Polystyrene with Nanoscale Bicontinuous Morphology.

The chloromethyl-functionalized polystyrene is the most commonly used ammonium cation precursor for making anion exchange resins (AER) and membranes (AEM). However, the chloromethylation of polystyrene or styrene involves highly toxic and carcinogenic raw materials (e.g., chloromethyl ether) and the resultant ammonium cation structural motif is not stable enough in alkaline media. Herein, we present a novel self-pored amine-functionalized polystyrene, which may provide a safe, convenient, and green process to make polystyrene-based AER and AEM. It is realized by hydrolysis of the copolymer obtained via random copolymerization of N-vinylformamide (NVF) with styrene (St). The composition and structure of the NVF-St copolymer could be controlled by monomeric ratio, and the copolymers with high NVF content could form bicontinuous morphology at sub-100 nm levels. Such bicontinuous morphology allows the copolymers to be swollen in water and self-pored by freeze-drying, yielding a large specific surface area. Thus, the copolymer exhibits high adsorption capacity (226 mg/g for bisphenol A). Further, the amine-functionalized polystyrene has all-carbon backbone and hydrophilic/hydrophobic microphase separation morphology. It can be quaternized to produce ammonium cations and would be an excellent precursor for making AEM and AER with good alkaline stability and smooth ion transport channels. Therefore, the present strategy may open a new pathway to develop porous alkaline stable AER and AEM without using metal catalysts, organic pore-forming agents, and carcinogenic raw materials.

Adsorption of Anthocyanins by Cation and Anion Exchange Resins with Aromatic and Aliphatic Polymer Matrices.

This study examines the mechanisms of adsorption of anthocyanins from model aqueous solutions at pH values of 3, 6, and 9 by ion-exchange resins making the main component of heterogeneous ion-exchange membranes. This is the first report demonstrating that the pH of the internal solution of a KU-2-8 aromatic cation-exchange resin is 2-3 units lower than the pH of the external bathing anthocyanin-containing solution, and the pH of the internal solution of some anion-exchange resins with an aromatic (AV-17-8, AV-17-2P) or aliphatic (EDE-10P) matrix is 2-4 units higher than the pH of the external solution. This pH shift is caused by the Donnan exclusion of hydroxyl ions (in the KU-2-8 resin) or protons (in the AV-17-8, AV-17-2P, and EDE-10P resins). The most significant pH shift is observed for the EDE-10P resin, which has the highest ion-exchange capacity causing the highest Donnan exclusion. Due to the pH shift, the electric charge of anthocyanin inside an ion-exchange resin differs from its charge in the external solution. At pH 6, the external solution contains uncharged anthocyanin molecules. However, in the AV-17-8 and AV-17-2P resins, the anthocyanins are present as singly charged anions, while in the EDE-10P resin, they are in the form of doubly charged anions. Due to the electrostatic interactions of these anions with the positively charged fixed groups of anion-exchange resins, the adsorption capacities of AV-17-8, AV-17-2P, and EDE-10P were higher than expected. It was established that the electrostatic interactions of anthocyanins with the charged fixed groups increase the adsorption capacity of the aromatic resin by a factor of 1.8-2.5 compared to the adsorption caused by the π-π (stacking) interactions. These results provide new insights into the fouling mechanism of ion-exchange materials by polyphenols; they can help develop strategies for membrane cleaning and for extracting anthocyanins from juices and wine using ion-exchange resins and membranes.

A universal method for the rapid isolation of all known classes of functional silencing small RNAs.

Diverse classes of silencing small (s)RNAs operate via ARGONAUTE-family proteins within RNA-induced-silencing-complexes (RISCs). Here, we have streamlined various embodiments of a Q-sepharose-based RISC-purification method that relies on conserved biochemical properties of all ARGONAUTEs. We show, in multiple benchmarking assays, that the resulting 15-min benchtop extraction procedure allows simultaneous purification of all known classes of RISC-associated sRNAs without prior knowledge of the samples-intrinsic ARGONAUTE repertoires. Optimized under a user-friendly format, the method - coined 'TraPR' for Trans-kingdom, rapid, affordable Purification of RISCs - operates irrespectively of the organism, tissue, cell type or bio-fluid of interest, and scales to minute amounts of input material. The method is highly suited for direct profiling of silencing sRNAs, with TraPR-generated sequencing libraries outperforming those obtained via gold-standard procedures that require immunoprecipitations and/or lengthy polyacrylamide gel-selection. TraPR considerably improves the quality and consistency of silencing sRNA sample preparation including from notoriously difficult-to-handle tissues/bio-fluids such as starchy storage roots or mammalian plasma, and regardless of RNA contaminants or RNA degradation status of samples.

Isolation and analysis of a non-protein low molecular weight thiol-mercurial adduct from human prostate lymph node cells (LNCaP).

Thiol compounds present in human malignant prostate cells (LNCaP) were investigated after reaction with a mercurial blocking reagent. After extracting the cellular glutathione and some other low molecular weight (LMW) thiols using trichloroacetic acid the resulting the protein precipitate was extracted with buffered 8 M urea containing 2-chloromercuri-4-nitrophenol in an equimolar amount to that of the thiol present. After removing the insoluble chromatin fraction the urea soluble labeled adducts formed were chromatographed on G15 Sephadex. Three yellow coloured (A410 nm) fractions were obtained; first, the excluded protein fraction containing 16.0 ± 4.1% of the applied label followed by an intermediate fraction containing 5.9 ± 1.2%. Finally a LMW fraction emerged which contained 77.2 ± 3.7% of the total label applied and this was further analyzed by column chromatography, first on an anion exchange column and then on a PhenylSepharose 6 column to give what appeared to be a single component. LC-MS analysis of this component gave a pattern of mercuri-clusters, formed on MS ionization showing possible parent ions at 704 or 588 m/z, the former indicating that a thiol fragment of molecular weight approximately 467 could be present. No fragments with a single sulfur adduct (a 369 m/z fragment) were observed The adduct was analyzed for cysteine and other amino acids, nucleic acid bases, ribose and deoxyribose sugars, selenium and phosphorus; all were negative leading to the conclusion that a new class of unknown LMW thiol is present concealed in the protein matrices of these cells.

The prevention of an anomalous chromatographic behavior and the resulting successful removal of viruses from monoclonal antibody with an asymmetric charge distribution by using a membrane adsorber in highly efficient, anion-exchange chromatography in flow-through mode.

Anion exchange (AEX) chromatography in the flow-through mode is a widely employed purification process for removal of process/product-related impurities and exogenous/endogenous viruses from monoclonal antibodies (mAbs). The pH of the mobile phase for AEX chromatography is typically set at half a unit below the isoelectric point (pI) of each mAb (i.e., pI - 0.5) or lower and, in combination with a low ionic strength, these conditions are usually satisfactory for both the recovery of the mAb and removal of impurities. However, we have recently encountered a tight binding of mAb1 to AEX resins under these standard chromatographic conditions. This anomalous adsorption behavior appears to be an effect of the asymmetric charge distribution on the surface of the mAb1. We found that mAb1 did not bind to the AEX resins if the mobile phase has a much lower pH and higher ionic strength, but those conditions would not allow adequate virus removal. We predicted that the use of membrane adsorbers might provide effective mAb1 purification, since the supporting matrix has a network structure that would be less susceptible to interactions with the asymmetric charge distribution on the protein surface. We tested the Natriflo HD-Q AEX membrane adsorber under standard chromatographic conditions and found that mAb1 flowed through the membrane adsorber, resulting in successful separation from murine leukemia virus. This AEX membrane adsorber is expected to be useful for process development because mAbs can be purified under similar standard chromatographic conditions regardless of their charge distributions.

Efficient removal of GenX (HFPO-DA) and other perfluorinated ether acids from drinking and recycled waters using anion exchange resins.

Carcinogenic GenX chemicals, heptafluoropropylene-oxide-dimer-acid (HFPO-DA), have been recently detected in surface, ground and recycled water sources worldwide. However, GenX removals under the influence of variable characteristics of the organic and inorganic compounds present in the natural water sources, have often been overlooked in scientific literature. This is critically important given that the ionic composition and characteristics of organic matter in natural waters are spatially and seasonally variable. A strongly basic anion exchange (IX) resin was used to remove GenX and two other perfluorinated ether acids (PFEAS) from natural surface and recycled water sources. Factors influencing the uptake behavior included the PFEAS concentrations, resin dosage, and background anion characteristics. The equivalent background compound was employed to evaluate the competitive uptake between natural organic matter (NOM), inorganic ions and PFEAS in natural water matrices. Experimental data were compared with different mathematical and physical models and it was depicted that approximately 4-6% of the initial NOM competed with PFEAS for active exchange sites. Further, IX was able to achieve complete PFEAS removal (Cfinal<10 ng/L) with simultaneous removal of>60% NOM and >80% inorganic ions. Results of this study indicate that IX exhibits great potential for PFEAS removal from natural drinking water sources.

A direct comparison between membrane adsorber and packed column chromatography performance.

The purpose of this work was to compare side by side the performance of packed bed and membrane chromatography adsorption processes for protein purification. The comparison was performed using anion exchange media with the same ligand immobilized on the adsorbing surface, namely the strong Q quaternary ammonium group, R-CH2-N+-(CH3)3, and bovine serum albumin (BSA) as a model protein. In addition, the stationary phase volume was held constant for each geometry (3 mL) and runs were executed using the same mobile phase superficial velocity. As expected, the packed bed column showed higher equilibrium binding of BSA at 66.9 mg/mL versus 43.04 mg/mL for the membrane adsorber. Dynamic binding capacities were also higher in the packed bed; for example, at 97.5 cm/h, a capacity of 62.8 mg/mL was measured for the packed bed versus 20.7 mg/mL for the membrane adsorber. The higher equilibrium and dynamic capacities of the packed bed are likely due to the higher surface area per unit volume of the resin. However, the maximum productivity for the membrane adsorber was 111 mg/(mL h), a value that was 3.3 times higher than the one of the packed column. The bed utilization - defined as the ratio of the dynamic binding capacity at 10% breakthrough to the saturation binding capacity - was also higher for the packed column at long residence times and lower at short residence times confirming the better performance of membrane chromatography at high flow rates.

Analytical Comparison of Methods for Extraction of Short Cell-Free DNA from Urine.

Urine cell-free DNA (cfDNA) is a valuable noninvasive biomarker for cancer mutation detection, infectious disease diagnosis (eg, tuberculosis), organ transplantation monitoring, and prenatal screening. Conventional silica DNA extraction does not efficiently capture urine cfDNA, which is dilute (ng/mL) and highly fragmented [30 to 100 nucleotides (nt)]. The clinical sensitivity of urine cfDNA detection increases with decreasing target length, motivating use of sample preparation methods designed for short fragments. We compared the analytical performance of two published protocols (Wizard resin/guanidinium thiocyanate and Q Sepharose), three commercial kits (Norgen, QIAamp, and MagMAX), and an in-house sequence-specific hybridization capture technique. Dependence on fragment length (25 to 150 nt), performance at low concentrations (10 copies/mL), tolerance to variable urine conditions, and susceptibility to PCR inhibition were characterized. Hybridization capture and Q Sepharose performed best overall (60% to 90% recovery), although Q Sepharose had reduced recovery (<10%) of the shortest 25-nt fragment. Wizard resin/guanidinium thiocyanate recovery was dependent on pH and background DNA concentration and was limited to <35%, even under optimal conditions. The Norgen kit led to consistent PCR inhibition but had high recovery of short fragments. The QIAamp and MagMAX kits had minimal recovery of fragments <150 and <80 nt, respectively. Urine cfDNA extraction methods differ widely in ability to capture short, dilute cfDNA in urine; using suboptimal methods may profoundly impair clinical results.

Selective removal of closely related clipped protein impurities using poly(ethylenimine)- grafted anion-exchange chromatography resin.

Proteolytic degradation is a serious problem that complicates downstream processing during production of recombinant therapeutic proteins. It can lead to decreased product yield, diminished biological activity, and suboptimal product quality. Proteolytic degradation or protein truncation is observed in various expression hosts and is mostly attributed to the activity of proteases released by host cells. Since these clipped proteins can impact pharmacokinetics and immunogenicity in addition to potency, they need to be appropriately controlled to ensure consistency of product quality and patient safety. A chromatography step for the selective removal of clipped proteins from an intact protein was developed in this study. Poly(ethylenimine)-grafted anion- exchange resins (PolyQUAT and PolyPEI) were evaluated and compared to traditional macroporous anion-exchange and tentacled anion-exchange resins. Isocratic retention experiments were conducted to determine the retention factors (k') and charge factors (Z) were determined through the classical stoichiometric displacement model. High selectivity in separation of closely related clipped proteins was obtained with the PolyQUAT resin. A robust design space was established for the PolyQUAT chromatography through Design-Of-Experiments (DoE) based process optimization. Results showed a product recovery of up to 63% with purity levels >99.0%. Approximately, one-log clearance of host cell protein and two-logs clearance of host cell DNA were also obtained. The newly developed PolyQUAT process was compared with an existing process and shown to be superior with respect to the number of process steps, process time, process yield, and product quality.

Retrospective Evaluation of Cycled Resin in Viral Clearance Studies-A Multiple Company Collaboration.

The BioPhorum Development Group Viral Clearance Workstream performed a collaborative retrospective analysis to evaluate packed bed chromatographic resin performance after repeated cycling for two commonly used chromatography steps in biopharmaceutical manufacturing: protein A and anion exchange. Key variables evaluated in the assessment included virus type, resin type, number of reuse cycles, and virus challenge. In this retrospective analysis of viral clearance data on naïve versus cycled resin, powered by the availability of a decade's worth of accumulated industry data, clearance capability was not negatively impacted by resin cycling. This finding is consistent with publications showing that surrogates for viral clearance capabilities could be employed in lieu of testing the viral clearance of cycled resins for protein A and anion exchange chromatography. The rigorous analysis of the retrospective data supports the view that viral clearance studies for cycled resins are not necessary provided that appropriate cleaning methods are applied during repeated use of the chromatography columns.LAY ABSTRACT: The manufacturing processes for biopharmaceutical products often include reusable chromatographic resins that remove process- and product-related impurities as well as potential contaminating viruses. Typically, chromatography resin is "cycled" through repeated steps of resin conditioning, product purification, and resin cleaning. The cycling approach has been evaluated in both small- and full-scale studies that show the performance parameters are maintained. The ability to remove virus is demonstrated separately in a focused small-scale virus-spiking study that is resource-intensive and costly. This paper is a retrospective review of industry data comparing virus removal by naïve and repeatedly cycled resins that summarizes the viral clearance impact of re-using protein A and anion exchange chromatography resins. The key variables evaluated in the assessment included virus type, resin type, number of cycles, and virus challenge. In this retrospective analysis, it was found that the viral clearance capability is not negatively impacted by resin cycling. This finding is consistent with other publications and supports the view that viral clearance studies for cycled resins are not necessary if appropriate cleaning methods are applied during the repeated use of the chromatography columns.Abbreviations: AAV-2, Adeno-associated virus; A-MuLV, Amphotropic murine leukemia virus; AEX, Anion-exchange chromatography; B/E, Bind and elute; BVDV, Bovine viral diarrhea virus; C.P.G., Controlled pore glass; DEAE, Diethylaminoethanol; EMCV, Encephalomyocarditis virus; FT, Flow through; HAV, Hepatitis A virus; HSV-1, Herpes simplex virus type 1; LOD, Limit of detection; LOQ, Limit of quantification; LRF, Log10 reduction factor; mAb, Monoclonal antibody; MVM, Minute virus of mice; NaOH, Sodium hydroxide; PA, Protein A; PPV, Porcine parvovirus; QA, Quaternary amine; QP, Quaternized polyethyleneimine; qPCR, Quantitative polymerase chain reaction; Reo3, Reovirus type 3; SuHV-1, Suid herpesvirus; SV40, Simian virus 40; X-MuLV, Xenotropic murine leukemia virus.

Adsorption and regeneration characteristics of phosphorus from sludge dewatering filtrate by magnetic anion exchange resin.

Removal and recovery of phosphorus (P) from sewage are essential for sustainable development of P resource. Based on the water quality determination of sludge dewatering filtrate from a wastewater treatment plant in Beijing, this study investigated the adsorption and regeneration characteristics of P by magnetic anion exchange resin (MAEX). The experiments showed that the P adsorption capacity of MAEX could reach a maximum of 2.74 mg/mL when initial P concentration was 25 mg/L and dosage of MAEX was 8 mL/L. The P adsorption on MAEX resin was suitable for large temperature range (283-323 K). However, the adsorption capacity was reduced in various degrees due to the interference of different anions (Br-, SO42-, Cl-, NO3-, HCO3-, CO32-) and organic compounds (bovine serum albumin, humic acid). Kinetics studies indicated that the P adsorption process followed the pseudo-second-order model. The MAEX resin had a rapid P adsorption rate and the P adsorption capacity at 30 min could reach 97.7-99.3% of qe. Increase of temperature was favorable to P adsorption on MAEX, and the adsorption isotherm data fitted to Langmuir model more than Freundlich model. Meanwhile, the thermodynamics parameters were calculated; it was shown that the adsorption process was an endothermic reaction. Desorption and regeneration experiments showed that NaHCO3 was a suitable regenerant, and the P adsorption capacity could reach 90.51% of the original capacity after 10 times of adsorption-desorption cycles; this indicated that MAEX resin has an excellent regeneration performance and thus has a very good application prospect of P removal and recovery. Fourier transform infrared spectroscopy (FTIR) analysis confirmed that ion exchange, charge attraction, and hydrogen bonding affected the removal of P by the MAEX resin. The vibrating sample magnetometer (VSM) analysis revealed that MAEX resin was a kind of soft magnetic materials with good magnetism.

Manipulating selectivity of covalently-bonded hyperbranched anion exchangers toward organic acids. Part I: Influence of primary amine substituents in the internal part of the functional layer.

Three covalently-bonded poly(styrene-divinylbenzene)-based (PS-DVB) hyperbranched anion exchangers prepared using primary amines with carboxylic, sulfonic or 2-hydroxyethyl substituents in the internal part of the functional layer were studied and compared for evaluating the effect of amines substituents on the chromatographic performance of the stationary phases. The hyperbranched coating was created on the surface of aminated PS-DVB substrate by repeating the modification cycles including alkylation with 1,4-butanediol diglycidyl ether (1,4-BDDGE) and amination with primary amine; glycine, taurine (2-aminoethanesulfonic acid) or ethanolamine were used in the first cycle, and 4 more cycles were conducted with methylamine (MA). The influence of the structure and type of primary amine used in the first cycle on the retention and selectivity toward inorganic anions, short-chain carboxylic acids, and polyphosphates was investigated using hydroxide eluent. The effect of temperature on the separation of organic acids was also studied for all stationary phases, which revealed the possibility to control and improve resolution for some pairs of analytes.