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Engineered particles adsorb biomolecules (e.g., proteins) when introduced in a biological medium to form a layer called a "corona". Coronas, in particular the protein corona, play an important role in determining the surface properties of particles and their targeting abilities. This study examines the influence of protein coronas on the targeting ability of layer-by-layer (LbL)-assembled polymer capsules and core-shell particles functionalized with monoclonal antibodies. Upon exposure of humanized A33 monoclonal antibody (huA33 mAb)-functionalized poly(methacrylic acid) (PMA) capsules or huA33 mAb-PMA particles to human serum, a total of 83 or 65 proteins were identified in the protein coronas, respectively. Human serum of varying concentrations altered the composition of the protein corona. The antibody-driven specific cell membrane binding was qualitatively and quantitatively assessed by flow cytometry and fluorescence microscopy in both the absence and presence of a protein corona. The findings show that although different protein coronas formed in human serum (at different concentrations), the targeting ability of both the huA33 mAb-functionalized PMA capsules and particles toward human colon cancer cells was retained, demonstrating no significant difference compared with capsules and particles in the absence of protein coronas: ∼70% and ∼90% A33-expressing cells were targeted by the huA33 mAb-PMA capsules and particles, respectively, in a mixed cell population. This result demonstrates that the formation of protein coronas did not significantly influence the targeting ability of antibody-functionalized LbL-polymer carriers, indicating that the surface functionality of engineered particles in the presence of protein coronas can be preserved.

We report the engineering of poly(ethylene glycol) (PEG) hydrogel particles using a mesoporous silica (MS) templating method via tuning the PEG molecular weight, particle size, and the presence or absence of the template and investigate the cell association and biodistribution of these particles. An ex vivo assay based on human whole blood that is more sensitive and relevant than traditional cell-line based assays for predicting in vivo circulation behavior is introduced. The association of MS@PEG particles (template present) with granulocytes and monocytes is higher compared with PEG particles (template absent). Increasing the PEG molecular weight (from 10 to 40 kDa) or decreasing the PEG particle size (from 1400 to 150 nm) reduces phagocytic blood cell association of the PEG particles. Mice biodistribution studies show that the PEG particles exhibit extended circulation times (>12 h) compared with the MS@PEG particles and that the retention of smaller PEG particles (150 nm) in blood, when compared with larger PEG particles (>400 nm), is increased at least 4-fold at 12 h after injection. Our findings highlight the influence of unique aspects of polymer hydrogel particles on biological interactions. The reported PEG hydrogel particles represent a new class of polymer carriers with potential biomedical applications.

The significance of a protein corona on nanoparticles in modulating particle properties and their biological interactions has been widely acknowledged. The protein corona is derived from proteins in biological fluids, many of which are glycosylated. To date, the glycans on the proteins have been largely overlooked in studies of nanoparticle-cell interactions. In this study, we demonstrate that glycosylation of the protein corona plays an important role in maintaining the colloidal stability of nanoparticles and influences nanoparticle-cell interactions. The removal of glycans from the protein corona enhances cell membrane adhesion and cell uptake of nanoparticles in comparison with the fully glycosylated form, resulting in the generation of a pro-inflammatory milieu by macrophages. This study highlights that the post-translational modification of proteins can significantly impact nanoparticle-cell interactions by modulating the protein corona properties.

Surface-confined ultra-thin polyrotaxane (PRX)-based films with tunable composition, surface topology and swelling characteristics were prepared by solid-state continuous assembly of polymers (ssCAP). The PRX-based films supported cell attachment, and their degradation in biological media could be tuned. This study provides a versatile nano-coating technology with potential applications in biomedicine, including tissue engineering and medical devices.

Studies of spherical nanoengineered drug delivery systems have suggested that particle size and mechanical properties are key determinants of in vivo behavior; however, for more complex structures, detailed analysis of correlations between in vitro characterization and in vivo disposition is lacking. Anisotropic materials in particular bear unknowns in terms of size tolerances for in vivo clearance and the impact of shape and rigidity. Herein, we employed cylindrical polymer brushes (CPBs) to answer questions related to the impact of size, length and rigidity on the in vivo behavior of PEGylated anisotropic structures, in particular their pharmacokinetics and biodistribution. The modular grafting assembly of CPBs allowed for the systematic tailoring of parameters such as aspect ratio or rigidity while keeping the overall chemical composition the same. CPBs with altered length were produced from polyinitiator backbones with different degrees of polymerization. The side chain grafts consisted of a random copolymer of poly[(ethylene glycol) methyl ether methacrylate] (PEGMA) and poly(glycidyl methacrylate) (PGMA), and rendered the CPBs water-soluble. The epoxy groups of PGMA were subsequently reacted with propargylamine to introduce alkyne groups, which in turn were used to attach radiolabels via copper(I)-catalyzed alkyne-azide cycloaddition (CuAAC). Radiolabeling allowed the pharmacokinetics of intravenously injected CPBs to be followed as well as their deposition into major organs post dosing to rats. To alter the rigidity of the CPBs, core-shell-structured CPBs with polycaprolactone (PCL) as a water-insoluble and crystalline core and PEGMA-co-PGMA as the hydrophilic shell were synthesized. This modular buildup of CPBs allowed their shape and rigidity to be altered, which in turn could be used to influence the in vivo circulation behavior of these anisotropic polymer particles. Increasing the aspect ratio or altering the rigidity of the CPBs led to reduced exposure

Particle stiffness is a design parameter that affects bionano interactions, including biodistribution kinetics and cellular processing. Herein, we develop soft polysaccharide (hyaluronic acid, HA) replica particles and capsules with tunable stiffness and sizes similar to human red blood cells (RBCs) via atom transfer radical polymerization-mediated continuous assembly of polymers (CAPATRP) and investigate their stiffness and deformability using colloidal-probe atomic force microscopy (CP-AFM) and a microfluidic blood capillary model, respectively. We demonstrate that HA replica particles and capsules with comparable nanoscale stiffness exhibit significantly different behaviors in a microfluidic blood capillary model. HA capsules behaved as RBCs, while HA replica particles had difficulty passing through the capillaries. These results (i) demonstrate how flow-based deformability measurements can be used to complement nanoscale stiffness measurements and (ii) provide important insight into the role of particle structure on the flow-based deformability of soft replica particles and capsules in a physiologically relevant microfluidic model.

Multilayer thin films have garnered intense scientific interest due to their potential application in diverse fields such as catalysis, optics, energy, membranes, and biomedicine. Here we review the current technologies for multilayer thin-film deposition using layer-by-layer assembly, and we discuss the different properties and applications arising from the technologies. We highlight five distinct routes of assembly—immersive, spin, spray, electromagnetic, and fluidic assembly—each of which offers material and processing advantages for assembling layer-by-layer films. Each technology encompasses numerous innovations for automating and improving layering, which is important for research and industrial applications. Furthermore, we discuss how judicious choice of the assembly technology enables the engineering of thin films with tailor-made physicochemical properties, such as distinct-layer stratification, controlled roughness, and highly ordered packing.

Protein coronas formed on engineered particles can alter their targeting ability as they enter biological environments. Here, we engineer polymer-coated silica particles and investigate the influence of protein coronas derived from various sources. The particles were functionalized with a small antibody-mimetic ligand (affibody), and their targeting ability to cancer cells in the presence of protein coronas was determined. Protein coronas derived from human serum showed a dramatic inhibition of specific particle-cell association (from ∼70 to ∼7%), whereas the most abundant protein in human serum-human serum albumin-enhanced the specific association of functionalized particles to SK-OV-3 human ovary cancer cells (to ∼90%). This study shows how protein coronas can both facilitate and impede targeting and provides key insights into the importance of challenging engineered particles with multicomponent biologically relevant environments.

It is demonstrated that metal-organic frameworks (MOFs) can be replicated in a biomimetic fashion from protein patterns. Bendable, fluorescent MOF patterns are formed with micrometer resolution under ambient conditions. Furthermore, this technique is used to grow MOF patterns from fingerprint residue in 30 s with high fidelity. This technique is not only relevant for crime-scene investigation, but also for biomedical applications.

Polymer microcapsules are of particular interest for applications including self-healing coatings, catalysis, bioreactions, sensing, and drug delivery. The primary way that polymer capsules can exhibit functionality relevant to these diverse fields is through the incorporation of functional cargo in the capsule cavity or wall. Diverse functional and therapeutic cargo can be loaded into polymer capsules with ease using polymer-stabilized calcium carbonate (CaCO3) particles. A variety of examples are demonstrated, including 15 types of cargo, yielding a toolbox with effectively 500+ variations. This process uses no harsh reagents and can take less than 30 min to prepare, load, coat, and form the hollow capsules. For these reasons, it is expected that the technique will play a crucial role across scientific studies in numerous fields.

Metal-phenolic coordination chemistry provides a simple and rapid way to fabricate ultrathin films. Here, we report a facile strategy for the preparation of low-fouling and pH-degradable metal-phenolic network (MPN) capsules using a synthetic polyphenol derivative, poly(ethylene glycol) (PEG)-polyphenol, as a building block. PEG-MPN capsules exhibit reduced nonspecific protein adsorption and cell association compared with tannic acid (TA)-MPN capsules. In addition, they show faster disassembly at a biologically relevant pH (5) than TA-MPN capsules (80% in 5 h vs 30% in 10 days). PEG-MPN capsules combine both the low fouling properties of PEG and the advantages of the MPN-driven assembly process (e.g., fast assembly and pH-degradability).

Protein particles are promising candidates for therapeutic delivery. In this study, we report a generalizable strategy to assemble a series of proteins into pH-cleavable protein particles that recover protein functionality after disassembly. Our strategy uses an acid-labile reversible cross-linker based on maleic anhydride chemistry, which allows the cross-linking of proteins and releases unmodified proteins upon cleavage, causing minimal loss of protein functionality. The protein particles can be rapidly disassembled at a mildly acidic pH (<6.5) and inside cells with negligible cytotoxicity. Furthermore, cleavage of the cross-linker led to above 97% recovery of enzymatic activity, as evidenced by using glucose oxidase. This facile and robust strategy to engineer pH-cleavable protein particles may provide a new platform for therapeutic protein delivery as well as for small molecule drug and nucleic acid delivery.

Antibody fusion to nonprotein materials such as contrast agents or radio-tracers, nano- or microparticles or small-molecule drugs is attracting major interest for molecular imaging and drug delivery. Nondirected bioconjugation techniques may impair antibody affinity, result in lower amounts of functional antibodies and generate multicomponent mixtures. We present a detailed protocol for the enzymatic bioconjugation of small recombinant antibodies to imaging particles, and we also describe the generation of and conjugation to a low-fouling capsule assembled for drug delivery from PEG and PVPON (poly(N-vinylpyrrolidone) by a layer-by-layer (LbL) technique. The single-chain variable fragment (scFv) is equipped with a short C-terminal LPETG tag and the fusion partners are functionalized with an N-terminal GGG nucleophilic group for sortase A conjugation. The LbL capsules are assembled through hydrogen bonding by depositing alkyne-modified poly(vinylpyrrolidone) and poly(methacrylic acid) layers on silica particles, followed by depositing alkyne-modified PEG. The generation of the antibodies and LbL capsules takes ∼1-2 weeks each. The conjugation and functional testing takes another 3-4 d.

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.

Layer-by-layer (LbL) assembly on nano- and microparticles is of interest for a range of applications, including catalysis, optics, sensors, and drug delivery. One current limitation is the standard use of manual, centrifugation-based (pellet/resuspension) methods to perform the layering steps, which can make scalable, highly controllable, and automatable production difficult to achieve. Here, we develop a fully flow-based technique using tangential flow filtration (TFF) for LbL assembly on particles. We demonstrate that multilayered particles and capsules with different sizes (from micrometers to submicrometers in diameter) can be assembled on different templates (e.g., silica and calcium carbonate) using several polymers (e.g., poly(allylamine hydrochloride), poly(styrenesulfonate), and poly(diallyldimethylammonium chloride)). The full system only contains fluidic components routinely used (and automated) in industry, such as pumps, tanks, valves, and tubing in addition to the TFF filter modules. Using the TFF LbL system, we also demonstrate the centrifugation-free assembly, including core dissolution, of drug-loaded capsules. The well-controlled, integrated, and automatable nature of the TFF LbL system provides scientific, engineering, and practical processing benefits, making it valuable for research environments and potentially useful for translating LbL assembled particles into diverse applications.

Smart poly(2-oxazoline) (POx)-based multifunctional polymer capsules that specifically target glycoprotein (GP) IIb/IIIa on the surface of activated platelets are degraded by the serine protease thrombin and release the urokinase plasminogen activator loaded into the polymer capsules, only in the area of acute thrombosis.

The chemical absorption of CO2 into a monoethanolamine solvent is currently the most widely accepted commercial approach to carbon dioxide capture. However, the subsequent desorption of CO2 from the solvents is extremely energy intensive. Alternative solvents are more energy efficient, but their slow reaction kinetics in the CO2 absorption step limits application. The use of a carbonic anhydrase (CA) enzyme as a reaction promoter can potentially overcome this obstacle. Native, engineered and artificial CA enzymes have been investigated for this application. Immobilization of the enzyme within the gas absorber or in a membrane format can increase enzyme stability and avoid thermal denaturation in the stripper. However, immobilization is only effective if the mass transfer of carbon dioxide through the liquid phase to reach the immobilization substrate does not become rate controlling. Further research should also consider the process economics of large-scale enzyme production and the long-term performance of the enzyme under real flue gas conditions.