Reactive Gelation Research Articles

Reactive Gelation Synthesis of Monodisperse Polymeric Capsules Using Droplet‐Based Microfluidics

AbstractA novel methodology, combining reactive gelation and droplet‐based microfluidics, is described for the synthesis of rigid, porous, and hollow polymeric nanoparticles (NPs). The precursors of such capsules are microfluidically generated latex droplets, with diameters tunable between 20 and 100 µm. The conversion of latex droplets to polymeric capsules involves two steps, namely self‐migration and gelation of the NPs toward the oil–water interface to form a solid‐like shell, and then postpolymerization to covalently fix the shell structure. The hollow structure of the capsules results from the interaction between negatively charged NPs inside the droplet and positive charges present on fluorosurfactant at the droplet interface. Significantly, the porosity and average pore size in the capsule shell can be controlled through variation of the initial NP concentration in the droplet. Based on the analysis of diffusion of fluorescent molecules of known size, it is shown that penetration of molecules into the internal volume of the capsules increases as the initial NP concentration in the droplet decreases, thus the porosity of the formed shell increases. This novel synthetic methodology defines a powerful tool in the generation of hollow capsules of controlled size and morphology, and has significant application in material sciences.

Macroporous Polymer⁻Protein Hybrid Materials for Antibody Purification by Combination of Reactive Gelation and Click-Chemistry.

Clickable core-shell nanoparticles based on poly(styrene-co-divinylbenzene-co-vinylbenzylazide) have been synthesized via emulsion polymerization. The 38 nm sized particles have been swollen by divinyl benzene (DVB) and 2,2’-azobis(2-methylpropionitrile) (AIBN) and subsequently processed under high shear rates in a Z-shaped microchannel giving macroporous microclusters (100 µm), through the reactive gelation process. The obtained clusters were post-functionalized by “click-chemistry” with propargyl-PEG-NHS-ester and propargylglicidyl ether, yielding epoxide or NHS-ester activated polymer supports for bioconjugation. Macroporous affinity materials for antibody capturing were produced by immobilizing recombinant Staphylococcus aureus protein A on the polymeric support. Coupling chemistry exploiting thiol-epoxide ring-opening reactions with cysteine-containing protein A revealed up to three times higher binding capacities compared to the protein without cysteine. Despite the lower binding capacities compared to commercial affinity phases, the produced polymer–protein hybrids can serve as stationary phases for immunoglobulin affinity chromatography as the materials revealed superior intra-particle mass transports.

Preparation of perfusive chromatographic materials via shear-induced reactive gelation

A simple method for producing highly porous materials suitable for chromatographic applications is discussed. Starting from a dispersion of polymer core-shell nanoparticles (latex), micrometer sized clusters (100 μm) are produced via shear-induced Reactive Gelation. Thanks to their fractal structure, these aggregates exhibit highly porous structures, with pore size distribution ranging from 0.1 to several micrometers. The effect of different properties of the primary nanoparticles on the qualities of the final products is also investigated. Particle architecture, namely the ratio between the hard, highly crosslinked core, and the soft, poorly crosslinked shell, turned out to be the most important parameter to be tuned in order to obtain highly porous and mechanically resistant clusters. The final materials can be easily slurry-packed into conventional chromatographic columns. In comparison to other commercial stationary phases, these materials show not only much lower pressure drops at very high flow rates (i.e. <0.2 bar/cm at 6 mL/min), but also HETP profiles independent of fluid velocity when measured with tracers of sizes comparable to typical bio-macromolecules. Moreover, these materials, while offering the key advantage of being in a slurry form and thus easily packable and scalable, have a behavior that closely resembles that of monoliths, in which convective flow contribution dominates.

Control of Pore Structure in Polymeric Monoliths Prepared from Colloidal Dispersions

AbstractReliable control of pore size distribution in porous materials is a key feature for addressing specific applications. The reactive gelation process represents a robust and efficient method to obtain mechanically stable monoliths with tunable pore size distribution. Primary polymer nanoparticles are destabilized and aggregated in a controlled way, forming a percolating gel. Afterward, this structure is hardened by a postpolymerization, carried out through heating. Different parameters play a major role in determining the final morphology of the monolith. In this work, the effect of primary particle architecture (i.e., core‐to‐shell ratio) and initial solid content of the latex is investigated, using two different sizes of nanoparticles. Actually, the first parameter affects the pores in the small range (0.01–1 µm) whereas the latter those in the larger one (1 µm to several micrometers), independently of the primary particle size. As a result, monoliths with very well‐defined pore size distributions are obtained.

Shear-Induced Reactive Gelation.

In this work, we describe a method for the production of porous polymer materials in the form of particles characterized by narrow pore size distribution using the principle of shear-induced reactive gelation. Poly(styrene-co-divinylbenzene) primary particles with diameter ranging from 80 to 200 nm are used as building blocks, which are assembled into fractal-like clusters when exposed to high shear rates generated in a microchannel. It was found that independent of the primary particle size, it is possible to modulate the internal structure of formed fractal-like aggregates having fractal dimension ranging from 2.4 to 2.7 by varying the residence time in the microchannel. Thermally induced postpolymerization was used to increase the mechanical resilience of such formed clusters. Primary particle interpenetration was observed by SEM and confirmed by light scattering resulting in an increase of fractal dimension. Nitrogen sorption measurements and mercury porosimetry confirmed formation of a porous material with surface area ranging from 20 to 40 m(2)/g characterized by porosity of 70% and narrow pore size distribution with an average diameter around 700 nm without the presence of any micropores. The strong perfusive character of the synthesized material was confirmed by the existence of a plateau of the height equivalent to a theoretical plate measured at high reduced velocities using a chromatographic column packed with the synthesized microclusters.

Application of polymeric macroporous supports for temperature-responsive chromatography of pharmaceuticals

A macroporous particulate support prepared previously by reactive gelation under shear and functionalized with poly(N-isopropylacrylamide), PNIPAM, brushes of variable length is applied for temperature-responsive chromatography, whereby temperature modulates hydrophobic interactions. Several different analytes, including small pharmaceuticals, peptides, proteins and monoclonal antibodies are employed. Contrary to the most commonly observed behavior in conventional chromatography, increasing retention is observed at elevated temperatures. Peak broadening is quantified using the peak standard deviation, which depends on both the polymer chain conformation and analyte adsorptivity. The favorable effect of grafted polymer thickness on retention becomes progressively less pronounced for thicker grafted PNIPAM layers. The effect of eluent composition on solute–sorbent interactions was investigated by introducing NaCl, methanol, dioxane and by varying the pH. Salt or organic solvent addition affects apart from the analytes solution properties, the hydrophobicity of the stationary phase itself. Frontal analyses performed at different temperatures to determine dynamic binding capacities, indicate small mass transfer resistances imposed by this novel packing material.

Perfusive ion-exchange chromatographic materials with high capacity

In this work, novel macro-porous chromatographic stationary phases, combining low mass transfer resistance and high binding capacity, were thoroughly characterized in terms of porosity, HETP, resolution and binding capacity. These new stationary phases exhibited better performance compared to commercially available materials, i.e. Poros 50HS and Fractogel EMD SO3 (M). With the technique of reactive gelation under shear, it is possible to produce particles with pores from 100nm to several microns, in which part of the flow can go through. This way, the mass transport inside the particles is significantly increased with perfusive flow faction values between 0.02 and 0.01. Despite the low pore surface area resulting from the large pore size, high binding capacity is obtained by functionalizing the pore surface with charged polymeric brushes resulting in a binding capacity in the range from 25 to 140mg/mL col. This, together with the high mass transfer, gives excellent resolution performance and dynamic binding capacity compared to other commercial materials even at high flow rates.

Macroporous polymer particles via reactive gelation under shear: effect of primary particle properties and operating parameters.

Reactive gelation under shear is a recently developed procedure toward rigid macroporous polymeric particles that does not require the use of any porogen. A comprehensive study of the effect of individual parameters on the resulting material characteristics is presented. Primary particle properties are found to be pivotal, namely, the primary particles size, cross-linking degree, and outer shell composition. Operating parameters also play a significant role; specifically, the effects of applied shear rate, salt feeding rate, swelling degree of primary particles, waiting time before postpolymerization, and postpolymerization temperature are investigated. By varying the operating conditions, the size, internal structure, as well as porosity of the fabricated microclusters may be controlled. The pores are invariably micrometer-sized, with pore size distributions exhibiting adjustable maxima. Thanks to the sequential character of the procedure, different parameters may be tuned individually at different stages along the preparation route, allowing thus for high versatility in the control of different properties of the final material.

Synthesis of macroporous polymer particles using reactive gelation under shear.

By combining elements from colloidal and polymer reaction engineering a new approach toward macroporous, mechanically robust polymer particles is presented, which does not require any porogenic additives. Specifically, aggregation and breakage in turbulent conditions of aggregates originating from fully destabilized primary latex particles is applied to produce compact, micrometer-sized clusters. Post-polymerization of monomer introduced initially to swell the primary particles is imparting mechanical rigidity and permanence to the internal structure. The resulting microclusters exhibit an internal porosity on the order of 70% and relatively broad pore size distribution, with exceptionally large pores, ranging from about 50 nm to 10 μm in diameter. These particulate microclusters, produced via reactive gelation under shear, are fractal objects with fractal dimension around 2.7, as opposed to the more open fractal structure of a monolith produced via stagnant reactive gelation, with fractal dimension of 1.9. Such macroporous particles are thought to be useful in applications requiring pores on the micrometer scale, e.g., in the chromatography of biomolecules or for packing beds perfusive to convective flow.

Strong cation exchange monoliths for HPLC by Reactive Gelation

Polymeric monolithic stationary phases for HPLC can be produced by Reactive Gelation. Unlike the conventional method of using porogens, such novel process consists of a number of separate steps, thus enabling a better control of the quality of the final material. A suspension of polymer nanoparticles in water is produced and subsequently swollen with hydrophobic monomers. The particles are then destabilised (usually by salt addition) to make them aggregate into a large percolating structure, the so-called monolith. Finally, the added monomer can then be polymerised to harden the structure. In this work, a polystyrene latex is used as the base material and functionalised by introduction of epoxide groups on the surface and subsequent reaction to sulphonic acid groups, yielding a SO3(-) density of 0.7 mmol/g dry material. Morphological investigations show 54% porosity made of 300 nm large pores. Van Deemter measurements of a large protein show no practical influence of diffusion limitations on the plate number. Finally, a preliminary separation of a test protein mixture is shown, demonstrating the potential of using ion-exchange chromatography on Reactive Gelation monoliths.

Preparation of macroporous methacrylate-based monoliths for chromatographic applications by the Reactive Gelation Process

Polymeric monoliths are a relatively new separation medium for chromatographic applications. The innovative approach to produce such monoliths, the Reactive Gelation Process, presented by Marti et al. [1] for polystyrene macroporous materials is applied to a methacrylate-based material. It is shown that it is possible to create a macroporous structure by Reactive Gelation also with this polymer even if the properties of the material are different. Besides the analysis of the material by SEM and BET, several chromatographic methods are used to analyze the material properties. The ISEC experiments showed a much smaller size exclusion effect than in conventional packed beds. The permeability of the material is comparable to a packed bed with 4.13 μ m particles. The column efficiency is not changing for increasing flow rates. Because of the high efficiency of the material, shorter columns are needed and therefore the comparatively low permeability is compensated. The monolith also exhibits a significant adsorption capacity for hydrophobic interaction, which makes it suitable for chromatographic purification processes.

PNIPAAM Grafted Polymeric Monoliths Synthesized by the Reactive Gelation Process and their Swelling/Deswelling Characteristics

The production of macroporous monoliths functionalized with a thermo-responsive polymer (PNIPAAM) is described. The surface functionalization was achieved by copolymerization of acrylic end capped atom transfer radical polymerization initiator (BPOEA) with divinylbenzene with or without styrene. Monoliths were generated by swelling them with styrene, BPOEA and divinylbenzene followed by gelation with salt and post polymerization. Subsequent grafting of these monoliths with PNIPAAM was achieved by atom transfer radical polymerization and their swelling deswelling characteristics quantified. The grafted monoliths provide a unique chromatographic stationary phase where adsorption/desorption can be driven by the use of temperature only.

Production of Polymeric Materials with Controlled Pore Structure: the “Reactive Gelation” Process

AbstractSummary: A new approach for the production of macroporous polymer materials is presented. Cross‐linked polystyrene particles are first produced by emulsion polymerization. This latex is then swollen by a further addition of monomer and it is successively destabilized, by addition of salt, in such a way that a controlled aggregation is achieved. The system is left aggregating until a gel is obtained. Although polymer gels have their own consistency, latex particles are kept together by weak interactions only, e.g. Van der Waals' forces. In order to impart enough mechanical resistance to the material, the residual monomer is polymerized, thus bridging the polymer particles with cross‐linked polymer. Accordingly, the process has been named “reactive gelation”. In this work, it is shown how it is possible to obtain an accurate control upon the pore structure of the material by properly tuning each step of the process. An application of the obtained material as stationary phase in a separation carried out by high performance liquid chromatography is presented.SEM picture of the macroporous material obtained by reactive gelation.magnified imageSEM picture of the macroporous material obtained by reactive gelation.