Application of mechanistic modelling in membrane and fiber chromatography for purification of biotherapeutics - A review.

Mechanistic modelling is a simulation tool which has been effectively applied in downstream bioprocessing to model resin chromatography. Membrane and fiber chromatography are newer approaches that offer higher rates of mass transfer and consequently higher flow rates and reduced processing times. This review describes the key considerations in the development of mechanistic models for these unit operations. Mass transfer is less complex than in resin columns, but internal housing volumes can make modelling difficult, particularly for laboratory-scale devices. Flow paths are often non-linear and the dead volume is often a larger fraction of the overall volume, which may require more complex hydrodynamic models to capture residence time distributions accurately. In this respect, the combination of computational fluid dynamics with appropriate protein binding models is emerging as an ideal approach.

The effect of calcium removal from skim milk by ion exchange on the properties of the ultrafiltration retentate.

New processes are needed to produce concentrated milk feedstocks with tailored calcium content, due to the direct link between calcium concentration and final product texture and functionality. Skim milk treatment with cation exchange resin 1% (w/v) or 2% (w/v) prior to ultrafiltration to a volumetric concentration factor (VCF) of 2.5 or 5 successfully decreased the calcium concentration by 20-30% and produced concentrates with solids content at ∼22-24 g 100 g-1 at a VCF of 5. Calcium reduction partially solubilized the casein micelles, increasing the concentration of soluble protein and individual caseins, leading to decreased turbidity but increased protein hydration and hydrophobicity. Decalcification (2% (w/v) resin treatment) reduced thermal stability, significantly decreasing the denaturation temperature of α-lactalbumin and ß-lactoglobulin in the milk by ∼3 °C and ∼1 °C respectively. Filtration was also altered, reducing permeation flux and the gel concentration and increased filtration time. When combined, calcium reduction and filtration altered functional properties including soluble calcium, soluble protein and sedimentable solids, with increased milk protein hydration also contributing to increased viscosity. This study provides a route to produce calcium-reduced milk concentrates with potential for use in retentate-based dairy products with tailored functionality.

Methanol Vapor Retards Aging of PIM-1 Thin Film Composite Membranes in Storage.

Physical aging of glassy polymers leads to a decrease in permeability over time when they are used in membranes. This hinders the industrial application of high free volume polymers, such as the archetypal polymer of intrinsic microporosity PIM-1, for membrane gas separation. In thin film composite (TFC) membranes, aging is much more rapid than in thicker self-standing membranes, as rearrangement within the thin active layer is relatively fast. Liquid alcohol treatment, which swells the membrane, is often used in the laboratory to rejuvenate aged self-standing membranes, but this is not easily applied on an industrial scale and is not suitable to refresh TFC membranes because of the risk of membrane delamination. In this work, it is demonstrated that a simple method of storage in an atmosphere of methanol vapor effectively retards physical aging of PIM-1 TFC membranes. The same method can also be utilized to refresh aged PIM-1 TFC membranes, and one-week methanol vapor storage is sufficient to recover most of the original CO2 permeance.

Industrial freezing and tempering for optimal functional properties in thawed Mozzarella cheese.

Mozzarella cheese was industrially frozen (-18 °C), stored for up to six months, tempered at 4 °C for one or three weeks and the structure and functionality compared to cheese stored at 4 °C and cheese aged at 4 °C for four weeks prior to freezing. When combined with ageing or tempering, the slow industrial freezing minimised changes to the protein network as detected by confocal microscopy and arrested proteolysis. Cheese functionality improved with three weeks of tempering, with properties similar to cheese refrigerated for one month, potentially due to increased proteolysis and protein rehydration. Frozen storage induced ß-sheet and ß-turn structures, as detected by S-FTIR microspectroscopy, with longer tempering leading to structural recovery in the cheese. This study indicates the proteolysis and functionality of frozen cheese can be optimised with tempering time. It also provides new insights into heat transfer during the industrial freezing and tempering of cheese.

Lactic Acid and Salt Separation Using Membrane Technology.

Acid whey is a by-product of cheese and yoghurt manufacture. The protein and lactose within acid whey can be recovered using nanofiltration and electrodialysis, but this leaves a waste stream that is a mixture of salts and lactic acid. To further add value to the acid whey treatment process, the possibility of recovering this lactic acid was investigated using either low energy reverse osmosis membranes or an electrodialysis process. Partial separation between lactic acid and potassium chloride was achieved at low applied pressures and feed pH in the reverse osmosis process, as a greater permeation of potassium chloride was observed under these conditions. Furthermore, lactic acid retention was enhanced by operating at lower temperature. Partial separation between lactic acid and potassium chloride was also achieved in the electrodialysis process. However, the observed losses in lactic acid increased with the addition of sodium chloride to the feed solution. This indicates that the separation becomes more challenging as the complexity of the feed solution increases. Neither process was able to achieve sufficient separation to avoid the use of further purification processes.

Recovery of Sevoflurane Anesthetic Gas Using an Organosilica Membrane in Conjunction with a Scavenging System.

Approximately 95% of the anesthetic gas administered to a patient is exhaled and ultimately released into the atmosphere. Most anesthetic gases have high global warming potential and so this approach adds significantly to the global greenhouse gas footprint. In this work, we develop a feasible means to capture such an anesthetic gas (sevoflurane) before it is released to the hospital scavenging system so that it is retained within the anesthetic circuit. Sevoflurane is retained using a microporous 1,2-bis(triethoxysilyl)ethane (BTESE) membrane prepared by a sol-gel method. The use of a ceramic membrane facilitates sanitization at high temperatures. A rapid thermal processing (RTP) technique is employed to reduce production time and to create a looser organosilica network, resulting in higher gas permeances, compared with the membrane synthesized from conventional thermal processing. The RTP membrane shows a slight decline in gas permeance when used with a dry mixture of CO2/N2/sevoflurane. This permeance falls again under 20% relative humidity feed conditions but the CO2/sevoflurane selectivity increases. The membrane performance shows little variation when the relative humidity is further increased. These promising results demonstrate that this microporous BTESE membrane has great potential for the recovery of sevoflurane in an anesthetic application.

Composite Membranes with Nanofibrous Cross-Hatched Supports for Reverse Osmosis Desalination.

A novel membrane structure composed of cross-hatched electrospun nanofibers is developed. We illustrate that this novel structure allows for much higher water permeability when used as a support for reverse osmosis thin-film composite membranes. Reinforcement and lamination of the aligned nanofibers generates mechanically robust structures that retain very high porosity and low tortuosity when applied to high pressure desalination operations. The cross-hatched nanofiber layers support the polyamide active layer firmly and reduce resistance to water flow due to the high porosity, low tortuosity, high mechanical strength, and minimal thickness of the structures. The nanofiber composite membrane gives a water flux significantly greater than when a traditional support layer is used, at 99 ± 5 m-2 h-1 with NaCl rejection of 98.7% at 15.5 bar.

The effect of pH on the fat and protein within cream cheese and their influence on textural and rheological properties.

The effect of variation in acid gel pH during cream cheese production was investigated. The gel microstructure was denser and cheese texture firmer, as the pH decreased from pH 5.0 to pH 4.3, despite the viscoelasticity of these gels remaining similar during heating. Protein hydration and secondary structure appeared to be key factors affecting both cheese microstructure and properties. Proteins within the matrix appeared to swell at pH 5.0, leading to a larger corpuscular structure; greater ß-turn structure was also observed by synchrotron-Fourier transform infrared (S-FTIR) microspectroscopy and the cheese was softer. A decrease in pH led to a denser microstructure with increased aggregated ß-sheet structure and a firmer cheese. The higher whey protein loss at low pH likely contributed to increased cheese hardness. In summary, controlling the pH of acid gel is important, as this parameter affects proteins in the cheese, their secondary structure and the resulting cream cheese.

The characterisation of Mozzarella cheese microstructure using high resolution synchrotron transmission and ATR-FTIR microspectroscopy.

Synchrotron Fourier transform infrared (S-FTIR) microspectroscopy allows the label-free examination of material microstructure but has not been widely applied to dairy products. Here, S-FTIR microspectroscopy was applied to observe the microstructure of Mozzarella cheese and assess the protein and lipid distribution within individual cheese blocks. High lipid and high protein areas were identified in transmission and attenuated total reflectance (ATR) analysis modes and the secondary structures of cheese proteins determined. Hierarchical cluster analysis and principal component analysis identified variation in random coil, water content, lipid carbonyl and methylene stretching across the sampled area. Similar spectral features were obtained in both analysis modes; spatial resolution was higher with ATR and small differences were noted, potentially as a result of differences in sample preparation. S-FTIR is a useful microscopy tool that can detect structural alterations that may affect product properties and may assist reverse engineering of a range of dairy products.

A comparison of multicomponent electrosorption in capacitive deionization and membrane capacitive deionization.

In this study, the desalination performance of Capacitive Deionization (CDI) and Membrane Capacitive Deionization (MCDI) was studied for a wide range of salt compositions. The comprehensive data collection for monovalent and divalent ions used in this work enabled us to understand better the competitive electrosorption of these ions both with and without ion-exchange membranes (IEMs). As expected, MCDI showed an enhanced salt adsorption and charge efficiency in comparison with CDI. However, the different electrosorption behavior of the former reveals that ion transport through the IEMs is a significant rate-controlling step in the desalination process. A sharper desorption peak is observed for divalent ions in MCDI, which can be attributed to a portion of these ions being temporarily stored within the IEMs, thus they are the first to leave the cell upon discharge. In addition to salt concentration, we monitored the pH of the effluent stream in CDI and MCDI and discuss the potential causes of these fluctuations. The dramatic pH change over one adsorption and desorption cycle in CDI (pH range of 3.5-10.5) can be problematic in a feed water containing components prone to scaling. The pH change, however, was much more limited in the case of MCDI for all salts.

Microstructure and physicochemical properties reveal differences between high moisture buffalo and bovine Mozzarella cheeses.

Mozzarella cheese is a classical dairy product but most research to date has focused on low moisture products. In this study, the microstructure and physicochemical properties of both laboratory and commercially produced high moisture buffalo Mozzarella cheeses were investigated and compared to high moisture bovine products. Buffalo and bovine Mozzarella cheeses were found to significantly differ in their microstructure, chemical composition, organic acid and proteolytic profiles but had similar hardness and meltability. The buffalo cheeses exhibited a significantly higher ratio of fat to protein and a microstructure containing larger fat patches and a less dense protein network. Liquid chromatography mass spectrometry detected the presence of only ß-casein variant A2 and a single ß-lactoglobulin variant in buffalo products compared to the presence of both ß-casein variants A1 and A2 and ß-lactoglobulin variants A and B in bovine cheese. These differences arise from the different milk composition and processing conditions. The differences in microstructure and physicochemical properties observed here offer a new approach to identify the sources of milk used in commercial cheese products.

The Role of Ion Exchange Membranes in Membrane Capacitive Deionisation.

Ion-exchange membranes (IEMs) are unique in combining the electrochemical properties of ion exchange resins and the permeability of a membrane. They are being used widely to treat industrial effluents, and in seawater and brackish water desalination. Membrane Capacitive Deionisation (MCDI) is an emerging, energy efficient technology for brackish water desalination in which these ion-exchange membranes act as selective gates allowing the transport of counter-ions toward carbon electrodes. This article provides a summary of recent developments in the preparation, characterization, and performance of ion exchange membranes in the MCDI field. In some parts of this review, the most relevant literature in the area of electrodialysis (ED) is also discussed to better elucidate the role of the ion exchange membranes. We conclude that more work is required to better define the desalination performance of the proposed novel materials and cell designs for MCDI in treating a wide range of feed waters. The extent of fouling, the development of cleaning strategies, and further techno-economic studies, will add value to this emerging technique.

Ultra-thin film composite mixed matrix membranes incorporating iron(III)-dopamine nanoparticles for CO2 separation.

Iron dopamine nanoparticles (FeDA NPs) are incorporated into a nanoscale thick polyethylene glycol (PEG) matrix for the first time, to form ultra-thin film composite mixed matrix membranes (UTFC-MMMs) via a recently developed continuous assembly of polymers (CAP) nanotechnology. The FeDA NPs are prepared by in situ nano-complexation between Fe(3+) and DA and have a particle size that can be varied from 3 to 74 nanometers by adjusting the molar ratio of DA to Fe(3+) ion. The cross-linked selective layer with sub 100 nanometer thickness is prepared by atom transfer radical polymerisation of a mixture of PEG macrocross-linkers and FeDA NPs on top of a highly permeable poly(dimethyl siloxane) (PDMS) prelayer, which is spin-coated onto a porous polyacrylonitrile (PAN) substrate. The incorporation of the FeDA NPs within the PEG-based selective layer is confirmed by XPS analysis. The UTFC-MMMs (thickness: ∼45 nm) formed present excellent gas separation performance with a CO2 permeance of ∼1200 GPU (1 GPU = 10(-6) cm(3) (STP) cm(-2) s(-1) cmHg(-1)) and an enhanced CO2/N2 selectivity of over 35, which is the best performance for UTFC membranes in the reported literature.

The dynamics of the biological membrane surrounding the buffalo milk fat globule investigated as a function of temperature.

The biological membrane surrounding fat globules in milk (the MFGM) is poorly understood, despite its importance in digestion and in determining the properties of fat globules. In this study, in situ structural investigations of buffalo MFGM were performed as a function of temperature (4-60°C), using confocal microscopy. We demonstrate that temperature and rate of temperature change affected the lipid domains formed in the MFGM with the lateral segregation (i) of high Tm lipids and cholesterol in a Lo phase for both TTm and (ii) of high Tm lipids in a gel phase for T

Crosslinked PEG and PEBAX Membranes for Concurrent Permeation of Water and Carbon Dioxide.

Membrane technology can be used for both post combustion carbon dioxide capture and acidic gas sweetening and dehydration of natural gas. These processes are especially suited for polymeric membranes with polyether functionality, because of the high affinity of this species for both H2O and CO2. Here, both crosslinked polyethylene glycol diacrylate and a polyether-polyamide block copolymer (PEBAX 2533(©)) are studied for their ability to separate CO2 from CH4 and N2 under single and mixed gas conditions, for both dry and wet feeds, as well as when 500 ppm H2S is present. The solubility of gases within these polymers is shown to be better correlated with the Lennard Jones well depth than with critical temperature. Under dry mixed gas conditions, CO2 permeability is reduced compared to the single gas measurement because of competitive sorption from CH4 or N2. However, selectivity for CO2 is retained in both polymers. The presence of water in the feed is observed to swell the PEG membrane resulting in a significant increase in CO2 permeability relative to the dry gas scenario. Importantly, the selectivity is again retained under wet feed gas conditions. The presence of H2S is observed to only slightly reduce CO2 permeability through both membranes.

Surface Engineering of Polypropylene Membranes with Carbonic Anhydrase-Loaded Mesoporous Silica Nanoparticles for Improved Carbon Dioxide Hydration.

Carbonic anhydrase (CA) is a native enzyme that facilitates the hydration of carbon dioxide into bicarbonate ions. This study reports the fabrication of thin films of active CA enzyme onto a porous membrane substrate using layer-by-layer (LbL) assembly. Deposition of multilayer films consisting of polyelectrolytes and CA was monitored by quartz crystal microgravimetry, while the enzymatic activity was assayed according to the rates of p-nitrophenylacetate (p-NPA) hydrolysis and CO2 hydration. The fabrication of the films onto a nonporous glass substrate showed CO2 hydration rates of 0.52 ± 0.09 µmol cm(-2) min(-1) per layer of bovine CA and 2.6 ± 0.7 µmol cm(-2) min(-1) per layer of a thermostable microbial CA. The fabrication of a multilayer film containing the microbial CA on a porous polypropylene membrane increased the hydration rate to 5.3 ± 0.8 µmol cm(-2) min(-1) per layer of microbial CA. The addition of mesoporous silica nanoparticles as a film layer prior to enzyme adsorption was found to increase the activity on the polypropylene membranes even further to a rate of 19 ± 4 µmol cm(-2) min(-1) per layer of microbial CA. The LbL treatment of these membranes increased the mass transfer resistance of the membrane but decreased the likelihood of membrane pore wetting. These results have potential application in the absorption of carbon dioxide from combustion flue gases into aqueous solvents using gas-liquid membrane contactors.

Effect of calcium chloride addition and draining pH on the microstructure and texture of full fat Cheddar cheese during ripening.

Calcium chloride is commonly added to cheese-milk to improve coagulum formation and to increase cheese yield but high concentrations of calcium ions can have adverse effects. In this study, confocal laser scanning microscopy and cryo-scanning electron microscopy were coupled with textural and chemical analyses to observe microstructural and biochemical changes that occur in cheese during ripening when calcium chloride is added or the draining pH altered. For the cheese prepared with no additional calcium at a draining pH of 6.0, the cheese porosity increased with ripening time and the number of protein vertices in the microscopy images declined, indicative of protein solubilisation. As the amount of CaCl2 added was increased, however, these changes became less significant. Our findings show that calcium chloride addition can be used, together with a lower draining pH, to alter the manufacturing process without significantly impacting on the quality of the mature cheese.

Chlorine resistant glutaraldehyde crosslinked polyelectrolyte multilayer membranes for desalination.

Crosslinked polyelectrolyte multilayer membranes are synthesized with salt rejection values approaching those of commercial desalination membranes, but with increased chlorine resistance. The membranes are fabricated directly onto porous commercial substrates. Subsequent crosslinking of the polycation layers with glutaraldehyde leads to NaCl rejections of up to 97%, while the incorporation of a highly sulfonated polysulfone polyanion leads to high chlorine resistance.

Influence of acoustic pressure and bubble sizes on the coalescence of two contacting bubbles in an acoustic field.

In this study, the coalescence time between two contacting sub-resonance size bubbles was measured experimentally under an acoustic pressure ranging from 10kPa to 120kPa, driven at a frequency of 22.4kHz. The coalescence time obtained under sonication was much longer compared to that calculated by the film drainage theory for a free bubble surface without surfactants. It was found that under the influence of an acoustic field, the coalescence time could be probabilistic in nature, exhibiting upper and lower limits of coalescence times which are prolonged when both the maximum surface approach velocity and secondary Bjerknes force increases. The size of the two contacting bubbles is also important. For a given acoustic pressure, bubbles having a larger average size and size difference were observed to exhibit longer coalescence times. This could be caused by the phase difference between the volume oscillations of the two bubbles, which in turn affects the minimum film thickness reached between the bubbles and the film drainage time. These results will have important implications for developing film drainage theory to account for the effect of bubble translational and volumetric oscillations, bubble surface fluctuations and microstreaming.

Experimental and theoretical analysis of secondary Bjerknes forces between two bubbles in a standing wave.

Bubbles in an acoustic field are affected by forces such as primary and secondary Bjerknes forces, which have been shown to be influenced by acoustic pressure, frequency, bubble size and separation distance between bubbles. However, such studies are predominantly theoretical, and are mostly focused on the sign reversal of the secondary Bjerknes force. This study provides experimental data on the effect of a range of bubble sizes (8-30 µm), distances (⩽0.2 mm), acoustic pressures (20-40 kPa) and frequencies (40-100 kHz) on the relative acceleration of two approaching bubbles. Under these conditions, only variations in the magnitude of the attractive force were observed. Using coupled equations of radial and translational motions, the acceleration and secondary Bjerknes force were calculated and compared to the experimental data. The variations in the magnitude of the secondary Bjerknes forces were explained by simulating bubble radius and coupled volume oscillation as a function of time.