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Fast-throughput and cost reduction of current purification platforms are becoming increasing requests during antibody manufacture. The macroporous-matrix absorbents have presented extensive potentiality in improving operational throughput during purification of macromolecule. And meanwhile the peptide ligand has become a promising alternative to recombinant protein ligands for cost reduction of chromatographic purification. Therefore, here we designed a functionalized microspheres resin with both macroporous matrix of polymerized glycidyl methacrylate and ethylene glycol dimethacrylate (PGMA-EDMA) and peptide ligand of hexapeptide (FYEILH). In order to circumvent the steric effect of peptides and amplify the binding sites on macroporous matrix, the peptide ligand was coupled on a liner PGMA polymer brushes grafted on microspheres. Comparing to the conventional agarose-matrix resin and the general peptide-grafted microspheres, the functionalized microspheres presented excellent permeability and high capacity to rapid loading hIgG by maintaining a stable level of dynamic binding capacity at fast flow rate above 110 column volume per hour (cv/h) and very short residence time below 0.5 min. Such functionalized microspheres provide a facile and broadly applicable strategy to develop the attractive candidate for rapid and cost-reduced purification of antibody., Competing Interests: Declaration of Competing Interest We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work. There is no professional or other personal interest of any nature or kind in any product, service and / or company that could be construed as influencing the position presented in, or the review of, the manuscript entitled., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Lipase (EC 3.1.1.3) is a versatile enzyme which has been widely used in ester-reaction industries. We have previously discovered that gigaporous polystyrene (PST) microspheres can be used as a novel immobilization carrier for lipase. In this work, a series of gigaporous microspheres with different densities of epoxy group including poly(glycidyl methacrylate) (PGMA) and poly(styrene-co-glycidyl methacrylate) [P(ST-GMA)] were evaluated as lipase immobilization carriers, which were also compared with gigaporous PST microspheres and the commercial immobilized lipase Novozym 435. Lipase immobilized in gigaporous PGMA microspheres showed the highest activity yield, reusability, and stability as well as the best affinity for the substrate. The characterizations of adsorption curves, the change of epoxy group amounts, and hydrophobic-hydrophilic properties of the microspheres were carried out to investigate the interaction between lipase molecules and carriers. It was found that covalent binding played a key role in improving the properties of lipase immobilized in gigaporous PGMA microspheres.

Great progress has been made in the preparation and application of multi-shelled hollow micro-/nanostructures during the past decade. However, the synthetic methodologies and potential applications of these novel and interesting materials have not been reviewed comprehensively in the literature. In the current review we first describe different synthetic methodologies for multi-shelled hollow micro-/nanostructures as well as their compositional and geometric manipulation and then review their applications in energy conversion and storage, sensors, photocatalysis, and drug delivery. The correlation between the geometric properties of multi-shelled hollow micro-/nanostructures and their specific performance in relevant applications are highlighted. These results demonstrate that the geometry has a direct impact on the properties and potential applications of such materials. Finally, the emerging challenges and future development of multi-shelled hollow micro-/nanostructures are further discussed.

Poly(DL-lactic-co-glycolic acid) (PLGA) microspheres have been widely prepared by many methods, including solvent evaporation, solvent extraction and the co-solvent method. However, very few studies have compared the properties of microspheres fabricated by these methods. This is partly because the broad size distribution of the resultant particles severely complicates the analysis and affects the reliability of the comparison. To this end, uniform-sized PLGA microspheres have been prepared by Shirasu porous glass premix membrane emulsification and used to encapsulate exenatide, a drug for treating Type 2 diabetes. Based on this technique, the influences on the properties of microspheres fabricated by the aforementioned three methods were intensively investigated, including in vitro release, degradation and pharmacology. We found that these microspheres presented totally different release behaviors in vitro and in vivo, but exhibited a similar trend of PLGA degradation. Moreover, the internal structural evolution visually demonstrated these release behaviors. We selected for further examination the microsphere prepared by solvent evaporation because of its constant release rate, and explored its pharmacodynamics, histology, etc., in more detail. This microsphere when injected once showed equivalent efficacy to that of twice-daily injections of exenatide with no inflammatory response., (Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Bio-waste cotton (Gossypium hirsutum L.) stalks were converted into succinic acid by simultaneous saccharification and fermentation (SSF) using Actinobacillus succinogenes 130Z. After 54 h SSF at 40 °C and pH 7.0, the production of succinic acid was 63 g/L, with 1.17 g/L/h productivity and 64% conversion yield. After SSF, a simple method for the decolorization and deproteinization of crude SSF broth was developed through adsorption tests of polystyrene (PSt) microspheres. Under optimized conditions (5% PSt loading (w/v), pH 4.0, 60 °C and adsorption time of 40 min), the ratios of decolorization, deproteinization and succinic acid loss ratios were 96.6, 84.5 and 4.1%, respectively. The method developed will provide a potential approach for large-scale production of succinic acid from the biomass waste., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

A novel sulfopropyl gigaporous polystyrene (SP-GP) microsphere enhancing the separation of poly(ethylene glycol)-protein (PEGylated protein) was first presented. The SP-GP microspheres were successfully prepared by introducing sulfopropyl groups into agarose-coated gigaporous polystyrene microspheres and used as chromatography media. Compared with a commercial medium, SP-GP microspheres exhibited improved column efficiency and reduced backpressure with increasing flow velocity, which could ensure its use in high-speed chromatography. Furthermore, a higher protein recovery and purity of the PEGylated protein could be obtained, even when SP-GP was applied at a flow velocity of 1224 cm h(-1). Additionally, the dynamic binding capacity (DBC) of SP-GP was significantly improved, which was higher than 10 mg mL(-1) medium even at a flow velocity of 306 cm h(-1). Further investigation using a laser scanning confocal microscope (LSCM) demonstrated that the static adsorption equilibrium of the PEGylated protein on SP-GP could be completed in 5 min, whereas a much longer period (ca. 60 min) was required for the commercial medium, indicating that the mass transfer of SP-GP was much faster with the gigaporous structure. All of these results strongly support that our developed SP-GP could serve as a promising cation exchange chromatography resin for high-speed separation, especially for biomolecules of high molecular weight., (Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.)

Rigid porous poly(glycidyl methacrylate-divinylbenzene) (P(GMA-DVB)) microspheres were synthesized through suspension polymerization with a mixture of isooctane and 4-methyl-2-pentonal as the porogen. The microspheres were intended to use as column packing materials for protein separation. However, irreversible adsorption of protein was found on the polymer microsphere. To circumvent the problem, polyethylene glycol (PEG) was coupled to the microspheres. The coupling reaction took place between the hydroxyl group of PEG and the epoxy group of the P(GMA-DVB) solid medium in the presence of boron trifluoride. The density of PEG immobilized onto the P(GMA-DVB) can be determined easily by saponification of modified microsphere firstly and then titration of glycerol-PEG. The effect of the cross-linker content of microsphere on the density of PEG immobilization was investigated. Molecular weight of PEG was found to influence the PEG-immobilization density, which subsequently affects the hydrophilicity of the modified P(GMA-DVB). Bovine serum albumin (BSA) and trypsin were used as model proteins to examine the adsorption and desorption properties of the modified P(GMA-DVB) microspheres. The results demonstrated that P(GMA-DVB) porous microsphere with 20% DVB and modified with PEG4000 showed excellent adsorption and desorption properties. Adsorption capacity of BSA on the modified microsphere attained to 51.6 mg/g microsphere, and BSA mass recovery and trypsin activity recovery was up to 97.6% and 98.7%, respectively. The modified microsphere was demonstrated to be a promising hydrophobic interaction chromatography material for purification of protein.

High resolution has been constantly pursued in both preparative and analytical chromatography. Chromatographic media are a key factor during the entire separation process. Tailor-made chromatographic media have gained more attention because of their adjustable structure appropriate for application. Uniform polysaccharide composite microspheres were prepared with a mixture of agarose and dextran solution by membrane emulsification technique for the first time. Their pore structure was deliberately regulated by adjusting both the polysaccharide composition and the molecular weight of dextran. Compared with pure agarose microspheres, polysaccharide composite microspheres had a higher separation resolution and their separation range was controllable. By increasing agarose concentration and decreasing dextran concentration at the same time during the preparation of composite microspheres, the mean pore size increased first and then decreased later, and also the pore size distribution became narrower. By increasing the molecular weight of dextran, the pores became smaller with a narrower pore size distribution. Microspheres with a composition of 10% agarose/2% dextran T40 or 8% agarose/4% dextran T150 showed a higher separation resolution for proteins within range of low molecular weight. Furthermore, the mechanical strength of this composite microsphere was improved by adjusting its composition. Atomic force microscope (AFM) results showed that pores were distributed evenly on both the surface and the inner part of microspheres, beneficial for the passage of biomolecules. These novel uniform polysaccharide composite microspheres have great potential for high-resolution bioseparation.ᅟ [ABSTRACT FROM AUTHOR]

Novel high-capacity Ni2+ immobilized metal ion affinity chromatographic media were prepared through the dextran-grafting process. Dextran was grafted to an allyl-activated agarose-based matrix followed by functionalization for the immobilized metal ion affinity chromatographic media. With elaborate regulation of the allylation degree, dextran was completely or partly grafted to agarose microspheres, namely, completely dextran-grafted agarose microspheres and partly dextran-grafted ones, respectively. Confocal laser scanning microscope results demonstrated that a good adjustment of dextran-grafting degree was achieved, and dextran was distributed uniformly in whole completely dextran-grafted microspheres, while just distributed around the outside of the partly dextran-grafted ones. Flow hydrodynamic properties were improved greatly after the dextran-grafting process, and the flow velocity increased by about 30% compared with that of a commercial chromatographic medium (Ni Sepharose FF). A significant improvement of protein binding performance was also achieved by the dextran-grafting process, and partly dextran-grafted Ni2+ chelating medium had a maximum binding capacity for His-tagged lactate dehydrogenase about 2.5 times higher than that of Ni Sepharose FF. The results indicated that this novel chromatographic medium is promising for applications in high-efficiency and large-scale protein purification. [ABSTRACT FROM AUTHOR]

Purpose: It is well known that primary emulsion (W/O) preparation process (by ultrasonication or homogenization) plays an important role in the properties of drug-loaded microspheres, such as encapsulation efficiency, release behavior and pharmacodynamics. However, its involved mechanism has not been intensively and systematically studied, partly because that broad size distribution of the resultant particles prepared by conventional preparation can greatly disturb the analysis and reliability of the results. Here, we focused on the relevant studies. Methods: In order to eliminate the disturbance caused by broad size distribution, uniform-sized exenatide-loaded poly(DL-lactic-co-glycolic acid) (PLGA) microspheres were prepared by Shirasu Porous Glass (SPG) premix membrane emulsification. The properties of microspheres whose W/O was formed by ultrasonication (UMS) and homogenization (HMS) were compared including in vitro release, pharmacology and so forth. Results: HMS exhibited fast release rate and hyperglycemic efficacy within first 14 days, but declined afterwards. Comparatively, UMS showed slower polymer degradation, more acidic microclimate pH (μpH) in vitro, and stable drug release with sustained efficacy during 1 month in vivo. Conclusions: HMS was desirable for the 2-week-sustained release in vivo, while UMS was more appropriate for the longer time release (about 1 month). These comparative researches can provide guidance for emulsion-microsphere preparation routs in pharmaceutics. [ABSTRACT FROM AUTHOR]

Two different polymers, poly(vinylbenzyl chloride) and poly(methyl methecrylate-2-hydroxyethyl methacrylate), were respectively employed as the major component of the particle material and amphiphilic polymer methoxy poly (ethylene glycol)-b-poly ( D, L-lactide) was employed as an essential additive. Gigaporous microspheres which possess flow-through pores which range from submicron to micron scale were prepared by two-step emulsification, emulsion ripening, and solvent extraction. Parameters on the final morphology were particularly investigated, including the polymer component, emulsion ripening time, and osmotic gradient. The formation mechanism of gigapores was also discussed with consideration of evolution behavior of the double emulsion system. The evolution process was proved crucial to control the final pore structure. Such kind of gigaporous microspheres could be further exploited as a promising chromatographic media for the purification of bioproducts. [ABSTRACT FROM AUTHOR]

A high cross-linking polystyrene(PSt)-based anion-exchange material with uniformly size, high ion exchange capacity, and high hydrophilicity was synthesized by a novel surface functionalization approach in this study. Uniformly sized PSt microspheres were prepared by the membrane emulsion polymerization strategy, and then modified by (1) conversing resid ual surface vinyl groups to epoxy groups followed by quaternization, and (2) decorating aromatic ring matrix including nitration, reduction and attachment of glycidyltrimethylammonium chloride. The 3-D morphology and porous features of microspheres were observed by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The surface of the modified PSt became roughness but the particle size remained same. Meanwhile, FT-IR spectra and laser scanning confocal microscope (LCSM) indicated that the modification groups had been successfully covalently coated onto the PSt microspheres. Modified PSt microspheres showed greatly improved hydrophilicity and biocompatibility with 0.387 mmol/mL ion exchange capacity (IEC). In the application evaluation procedure, exenatide can be purified from 42.9% (peptide crudes) to 88.6% by modified PSt column with 97.1% recovery yield. This modified PSt microspheres had a large potential in application for efficient separation of peptides. [ABSTRACT FROM AUTHOR]

Highlights: [•] A novel GSH affinity medium was prepared based on macroporous rigid microspheres. [•] GSH-AP column had less backpressure and plate height than those of GSH-FF column. [•] GSH-AP column had higher dynamic binding capacity at high flow velocity. [•] GSH-AP medium was promising for high-speed affinity chromatography. [ABSTRACT FROM AUTHOR]

Highlights: [•] Uniform-sized PLGA microspheres with large size (around 20μm) were prepared by SPG premix membrane emulsion technique, which has not been reported. [•] The in vitro release of exenatide-loaded PLGA microspheres prepared by this technique was improved by optimization of process parameters. [•] Bio-stability of exenatide inside microspheres was not influenced by the preparation process. [•] This technique was extended to prepare microspheres with size range from hundreds of nanometers to tens of microns. [ABSTRACT FROM AUTHOR]

Abstract: Protein denaturation and nonspecific adsorption on polymer media as a chromatographic support have been a problem which needs to be overcome. Macroporous poly(glycidyl methacrylate–divinylbezene) (PGMA–DVB) microspheres prepared in this study were firstly covalently coated with dextran through a three-step method. The dextran was firstly adsorbed onto the microspheres and then covalently bound to the PGMA–DVB microsphere through ether bonds which were formed by hydroxyl group reacting with epoxy group at the presence of 4-(Dimethylamino) pyridine. Finally, the coating dextran layer was crosslinked by ethylene glycol diglycidyl ether to form the continuous network coating. The coated microspheres were characterized by Fourier transform infrared spectra, scanning electron microscope, mercury porosimetry measurements, laser scanning confocal microscope, and protein adsorption experiments. Results showed that PGMA–DVB microspheres coated with dextran successfully maintained the macroporous structure and high permeability. The backpressure was only 1.69MPa at a high flow rate of 2891cm/h. Consequently, the hydrophilicity and biocompatibility of modified microspheres were greatly improved, and the contact angle decreased from 184° to 13°, and nonspecific adsorption of proteins was decreased to little or none. The clad dextran coating with large amounts of hydroxyl group was easily derived to be various functional groups. The derived media have great potential applications in rapid protein chromatography. [Copyright &y& Elsevier]

Abstract: In this article, a simple way to control pore size in monodispersed poly(N-Isopropyl acrylamide-co-acrylic acid) [P(NIPAM-co-AAc)] microspheres just by changing the acrylic acid wt.% in the formula was developed. The monodispersed P(NIPAM-co-AAc) microspheres with functionalized carboxyl groups were successfully prepared by Shirasu porous glass (SPG) membrane emulsification technique at room temperature. The microspheres were dried by supercritical drying (SCD), lyophilization and vacuum drying, separately. It was observed that SCD was an effective drying method to maintain the morphologies of the microspheres. Scanning electron microscopy (SEM), confocal laser scanning microscope (CLSM), and porosity analyzer results showed that the P(NIPAM-co-AAc) microspheres contained abundant slit-like mesoporous structure with pore size from 2nm to 50nm. The increase of AAc wt.% would increase the percentage of mesopores and micropores in the microspheres. P(NIPAM-co-AAc) microspheres with specific pore size can be obtained by simply changing the acrylic acid wt.% in the formula. Such temperature-/pH-ensitive microspheres with controllable pore size were expected to meet various requirements in different bioapplications, such as drug delivery and enzyme immobilization. [Copyright &y& Elsevier]