Synthesis of Polymer Brushes on Tannic Acid-Coated Copper Particles and Surface Co-Assembly.

The synthesis of polymer brushes on inorganic particles is an effective approach to surface modification. The polymer brushes on the surface endow the substrates with new surface properties. However, the lack of functional groups and the difficulty of surface modification have made it difficult to develop an effective method for the synthesis of polymer brushes on metal surfaces. Herein, a simple and versatile strategy for synthesizing polymer brushes on copper particles is reported. Tannic acid (TA) molecules are adsorbed onto the surfaces of copper particles, forming TA coatings. Quaternized poly(2-(dimethylamino)ethyl methacrylate)-block-polystyrene (qPDMAEMA-b-PS) block copolymer (BCP) chains are grafted on the TA coatings through hydrogen bonding and electrostatic interaction, and PS brushes are grafted on the copper particles. The effects of TA concentration on the adsorption of TA and PS brush synthesis are discussed. The PS brushes are able to form surface nanostructures on the copper particles through co-assembly with PDMAEMA-b-PS BCP chains. The effect of BCP concentration on the surface nanostructures is investigated. It is reasonable to expect that polymer brushes and surface nanostructures can be synthesized on different metal surfaces by using the TA-coating approach reported in this paper.

Synergistic Effects of the Superhydrophilic and Superhydrophobic Components on the Antifreezing Performances of Latex Particles and Anti-Icing Properties of Latex Films.

The development of new materials for antifreezing and anti-icing applications is a big challenge in industry and academic area. Inspired by the antifreeze proteins, latex particles with superhydrophilic zwitterionic shells and superhydrophobic cores are synthesized by reversible addition-fragmentation chain transfer emulsion polymerization, and the applications of the latex particles in antifreezing and anti-icing applications are investigated. In antifreezing study, the critical aggregate temperature (CAT) of the latex particles decreases, and the separation of the melting and freezing temperature of ice increases with the particle concentration. Enzyme molecules can be cryopreserved in the particle solution, and their bioactivities are well maintained. Latex particles are casted into latex films with dynamic surfaces. Anti-icing performances, including antifrosting properties, freezing delay time, and ice adhesion strengths, are studied; and the water-treated latex films present stronger anti-icing properties than other films, due to the synergistic effects of the superhydrophilic and superhydrophobic components. In addition, latex particles with zwitterionic shells and poly(n-butyl methacrylate) cores, and latex particles with small molecular surfactant on the surfaces are synthesized. The antifreezing performances of the latex particles and anti-icing properties of the latex films are compared.

Pore-blocking steric mass-action model for adsorption of bioparticles.

The steric mass-action (SMA) model has been widely reported to describe the adsorption of proteins in different types of chromatographic adsorbents. Here in the present work, a pore-blocking steric mass-action model (PB-SMA) was developed for the adsorption of large-size bioparticles, which usually exhibit the unique pore-blocking characteristic on the adsorbent and thus lead to a fraction of ligands in the deep channels physically inaccessible to bioparticles adsorption, instead of being shielded due to steric hindrance by adsorbed bioparticles. This unique phenomenon was taken into account by introducing an additional parameter, Lin, which is defined as the inaccessible ligand densities in the physically blocked pore area, into the PB-SMA model. This fraction of ligand densities (Lin) will be deducted from the total ligand (Lt) for model development, thus the steric factor (σ) in the proposed PB-SMA will reflect the steric shielding effect on binding sites by adsorbed bioparticles more accurately than the conventional SMA model, which assumes that all ligands on the adsorbent have the same accessibility to the bioparticles. Based on a series of model assumptions, a PB-SMA model was firstly developed for inactivated foot-and-mouth disease virus (iFMDV) adsorption on immobilized metal affinity chromatography (IMAC) adsorbents. Model parameters for static adsorption including equilibrium constant (K), characteristic number of binding sites (n), and steric factor (σ) were determined. Compared with those derived from the conventional SMA model, the σ values derived from the PB-SMA model were dozens of times smaller and much closer to the theoretical maximum number of ligands shielded by a single adsorbed iFMDV, indicating the modified model was more accurate for bioparticles adsorption. The applicability of the PB-SMA model was further validated by the adsorption of hepatitis B surface antigen virus-like particles (HBsAg VLPs) on an ion exchange adsorbent with reasonably improved accuracy. Thus, it is considered that the PB-SMA model would be more accurate in describing the adsorption of bioparticles on different types of chromatographic adsorbents.

Polymer Brushes and Surface Nanostructures: Molecular Design, Precise Synthesis, and Self-Assembly.

For over two decades, polymer brushes have found wide applications in industry and scientific research. Now, polymer brush research has been a significant research focus in the community of polymer science. In this review paper, we give an introduction to the synthesis, self-assembly, and applications of one-dimensional (1D) polymer brushes on polymer backbones, two-dimensional (2D) polymer brushes on flat surfaces, and three-dimensional (3D) polymer brushes on spherical particles. Examples of the synthesis of polymer brushes on different substrates are provided. Studies on the formation of the surface nanostructures on solid surfaces are also reviewed in this article. Multicomponent polymer brushes on solid surfaces are able to self-assemble into surface micelles (s-micelles). If the s-micelles are linked to the substrates through cleavable linkages, the s-micelles can be cleaved from the substrates, and the cleaved s-micelles are able to self-assemble into hierarchical structures. The formation of the surface nanostructures by coassembly of polymer brushes and "free" polymer chains (coassembly approach) or polymerization-induced surface self-assembly approach, is discussed. The applications of the polymer brushes in colloid and biomedical science are summarized. Finally, perspectives on the development of polymer brushes are offered in this article.

Polymer-Protein Nanovaccine Synthesized via Reactive Self-Assembly with Potential Application in Cancer Immunotherapy: Physicochemical and Biological Characterization In Vitro and In Vivo.

Nanovaccines composed of polymeric nanocarriers and protein-based antigens have attracted much attention in recent years because of their enormous potential in the prevention and treatment of diseases such as viral infections and cancer. While surface-conjugated protein antigens are known to be more immunoactive than encapsulated antigens, current surface conjugation methods often result in low and insufficient protein loading. Herein, reactive self-assembly is used to prepare nanovaccine from poly(ε-caprolactone) (PCL) and ovalbumin (OVA)-a model antigen. A rapid thiol-disulfide exchange reaction between PCL with pendant pyridyl disulfide groups and thiolated OVA results in the formation of nanoparticles with narrow size distribution. High OVA loading (≈70-80 wt%) is achieved, and the native secondary structure of OVA is preserved. Compared to free OVA, the nanovaccine is much superior in enhancing antigen uptake by bone marrow-derived dendritic cells (BMDCs), promoting BMDC maturation and antigen presentation via the MHC I pathway, persisting at the injection site and draining lymph nodes, activating both Th1 and Th2 T cell immunity, and ultimately, resisting tumor challenge in mice. This is the first demonstration of reactive self-assembly for the construction of a polymer-protein nanovaccine with clear potential in advancing cancer immunotherapy.

Synthesis of Cleavable Polymers via Oxidation of Thioether Moieties and Thiol Click Reactions: A Click-Declick Strategy.

In the past decades click chemistries including thiol chemistries have found wide applications in the synthesis of well-defined polymers. In this research, a click-declick strategy based on the oxidation of heteroaromatic thioethers and the substitution reactions between the oxidized groups and thiols, is proposed for the synthesis of the cleavable polymers. In proof-of-concept experiments, block copolymers (BCPs) and star-like polymers are synthesized by thiol-phenylsulfone substitution reactions, and heteroaromatic thioethers are produced at the junction points of the BCP chains or on the crosslinking sites of the star-like polymer. The thioethers can be oxidized to heteroaromatic sulfoxides or sulfones, depending on the oxidization condition. It is demonstrated that both sulfoxides or sulfones can have base catalyzed nucleophilic substitution reactions with thiols, leading to the cleavage of the polymers.

Polymerization-Induced Proteinosome Formation Initiated by Artificial Cells.

Cellular communication is essential for living cells to coordinate the individual cellular responses and make collective behaviors. In the past decade, the communications between artificial cells have aroused great interest due to the potential applications of the structures in bioscience and biotechnology. To mimic the cellular communication, artificial cell assisted synthesis of proteinosomes was studied in this research. Multienzyme proteinosomes with glucose oxidase (GOx) and horseradish peroxidase (HRP) decorated on the membranes were synthesized by the thermally triggered self-assembly approach. Free radicals produced in a cascade reaction taking place on the surfaces of the multienzyme proteinosomes initiated reversible addition-fragmentation chain transfer (RAFT) polymerization of NIPAM at a temperature above LCST of PNIPAM in the presence of bovine serum albumin (BSA) or alcohol dehydrogenase (ADH)/acetaldehyde dehydrogenase (ALDH), and daughter proteinosomes with BSA or ADH/ALDH on the surfaces were fabricated. The structures of the GOx/HRP initiator proteinosomes, and the synthesized daughter proteinosomes were characterized with transmission electron microscopy, atomic force microscopy, fluorescence microscopy, dynamic light scattering, and micro-DSC. Enzyme activity assays demonstrate the high bioactivities of the enzymes on the surfaces of the initiator and the synthesized daughter proteinosomes.

Visible light-promoted transition metal-free direct C3-carbamoylation of 2H-Indazoles.

We reported a general transition metal-free transformation to access C3-carbamoylated 2H-indazoles via visible light-induced oxidative decarboxylation coupling, in the presence of oxamic acids as the coupling sources, 4CzIPN as the photocatalyst, and Cs2CO3 as the base. The great application potential of this mild condition is highlighted by the late-stage modification of drugs, N-terminal modification of peptides, and the good antitumor activity of the novel desired product.

Surface Coassembly of Binary Mixed Polymer Brushes and Linear Block Copolymer Chains.

Binary mixed polymer brushes (BMPBs) are two different homopolymer chains that are covalently anchored to the solid surfaces at high grafting densities. One feature of the BMPBs is the unique ability to make surface phase separation under external stimuli. In this research, we demonstrate that different surface nanostructures can be fabricated by surface coassembly of BMPBs and free block copolymer (BCP) chains. Polystyrene/poly(2-(dimethylamino)ethyl methacrylate) (PS/PDMAEMA) BMPBs on silica particles (PS-PDMAEMA-SiO2) are synthesized by a two-step "grafting to" approach. PDMAEMA-b-PS block copolymer (BCP) chains and PS-PDMAEMA-SiO2 make surface self-assembly and a variety of surface nanostructures are formed in methanol. The grafting densities of PS and PDMAEMA brushes, solvent, and the BCP structures all exert significant influences on the surface morphology. With an increase in PDMAEMA grafting density, the surface structures change from perforated layers, to rods, and to spherical surface micelles (s-micelles). The PS grafting density also exerts an effect on the formation of the surface nanostructures. At low PS grafting density, sparsely distributed s-micelles are produced, and at high density, densely distributed s-micelles are observed. Based on transmission electron microscopy and scanning electron microscopy results, a surface phase diagram is constructed, which provides a guide to the surface morphology control.

Polymer brush-based nanostructures: from surface self-assembly to surface co-assembly.

Surface structures play an important role in the practical applications of materials. The synthesis of polymer brushes on a solid surface has emerged as an effective tool for tuning surface properties. The fabrication of polymer brush-based surface nanostructures has greatly facilitated the development of materials with unique surface properties. In this review article, synthetic methods used in the synthesis of polymer brushes, and self-assembly approaches applied in the fabrication of surface nanostructures including self-assembly of polymer brushes, co-assembly of polymer brushes and "free" block copolymer chains, and polymerization induced surface self-assembly, are reviewed. It is demonstrated that polymer brush-based surface nanostructures, including spherical surface micelles, wormlike surface structures, layered structures and surface vesicles, can be fabricated. Meanwhile, the challenges in the synthesis and applications of the surface nanostructures are discussed. This review is expected to be helpful for understanding the principles, methods and applications of polymer brush-based surface nanostructures.

Ca2+-Chelation-Induced Fabrication of Multistimuli-Responsive Charged Nanogels from Phospholipid-Polymer Conjugates and Use for Drug/Protein Loading.

Thermoresponsive phospholipid-poly(N-isopropylacrylamide) (PL-PNIPAM) conjugates were synthesized via reversible addition fragmentation chain transfer polymerization mediated by a phospholipid-modified trithiocarbonate. Temperature triggered the micellization of the PL-PNIPAM conjugate to form phosphate group-decorated micelles in the aqueous solution. Driven by the chelation of phospholipids and Ca2+, the PL-PNIPAM conjugate and Ca2+ ions formed size-tunable nanoclusters at a temperature beyond the lower critical solution temperature. To fabricate cross-linked nanogels, NIPAM was copolymerized with N-succinimidyl acrylate (NSA) to obtain the PL-P(NIPAM-co-NSA) conjugate bearing pendent cross-linkable functionalities. Subsequently, the size-controllable nanogels containing disulfide linkages were generated at 37 °C by cross-linking the PL-P(NIPAM-co-NSA)/Ca2+ nanoclusters with cystamine through modulation of Ca2+ concentrations. These negatively charged nanogels demonstrate temperature/pH/reduction triple responsiveness. The nanogels can be efficiently loaded with doxorubicin (DOX) and proteins with various isoelectric points. The DOX-loaded nanogels exhibited a temperature/pH/reduction triple-responsive release profile. The immobilized RNase A, BSA, and GOx retained the protein bioactivity. The release of RNase A-loaded nanogels possesses a temperature-responsive profile. The immobilization of Lys and cytochrome C in nanogels inhibited protein bioactivity. However, the addition of NaCl triggered the recovery of bioactivity. These multistimuli-responsive nanogels can provide a versatile platform applicable in biotechnology and drug/protein delivery.

Synthesis and modification of polymers by thiol-phenylsulfone substitution reaction.

Thiol chemistry is a type of highly efficient chemical reaction between thiols and functional groups. During the past two decades, thiol chemistry has been widely applied in the synthesis and modification of polymers. With the rapid development of polymer chemistry and materials science, more thiol click reactions, which can be efficiently performed under mild conditions, are required. In this research, the synthesis and modification of polymers by thiol-phenylsulfone substitution reactions are reported. A monomer containing two phenylsulfonyl groups is synthesized and the monomer is reacted with bisthiols under mild conditions, leading to the synthesis of novel polymers. Size exclusion chromatography, 1H NMR and differential scanning calorimetry results demonstrate the step-growth polymerization of the monomer. A combination of thiol-phenylsulfone and thiol-disulfide reactions are used in the post-polymerization modification.

Enzyme-catalyzed cascade reactions on multienzyme proteinosomes.

In this research, to mimic the structures and the functionalities of the organelles in living cells multienzyme proteinosomes with ß-galactosidase (ß-gal), glucose oxidase (GOx) and horseradish peroxidase (HRP) on the surfaces are fabricated by hydrophobic-interaction induced self-assembly approach. To investigate the mechanism of the formation of proteinosomes, poly(di(ethylene glycol) methyl ether methacrylate) (PDEGMA) and bovine serum albumin are employed in a model system and the study demonstrates that the hydrophobic interaction between the dehydrated polymer chains and the hydrophobic patches on the proteins plays a key role in the fabrication of the proteinosomes. Based on the model system, multienzyme proteinosomes with ß-gal, GOx and HRP on the surfaces are fabricated through hydrophobic interaction between PDEGMA and enzyme molecules. Enzyme-catalyzed cascade reactions are performed on the surfaces of the proteinosomes, and the immobilized enzymes show higher bioactivities than the "free" enzymes, due to the direct transfer of the product as a substrate from one enzyme molecule to another. This research provides a unique method for the synthesis of multienzyme proteinosomes with improved bioactivities, and the biofunctional structures will find promising applications in medical and biological science.

Fabrication of PεCL-AuNP-BSA core-shell-corona nanoparticles for flexible spatiotemporal drug delivery and SERS detection.

Nanoparticles with protein coronae can be used as promising multifunctional platforms for nanomedicine due to the possibility of performing surface functionalization on protein molecules and the achievement of biomedical properties. In this research, nanoparticles (NPs) with poly(ε-caprolactone) (PεCL) cores, gold NP (AuNP) shells and BSA coronae were fabricated by a self-assembly approach. The hydrophobic PεCL cores were used to encapsulate curcumin (CUR), the AuNP shells were decorated with a Raman probe, and the protein molecules in the coronae were functionalized with folic acid (FA). The self-assembly behaviors, drug delivery and the surface-enhanced Raman scattering (SERS) effect of the hybrid NPs were investigated in this research. The sizes of the core-shell-corona NPs (CSCNPs) are dependent on the initial concentrations of PεCL and AuNPs. The CUR in CSCNPs show enzyme-triggered release properties. The added lipase or trypsin can facilitate the CUR release from the hybrid NPs. The functionalization of CSCNPs with FA can significantly improve the internalization of NPs into 4T1 tumor cells due to the overexpressed folate receptors on the cells. In addition, the SERS effect of CSCNPs can be achieved when the AuNPs are decorated with 2-naphthalenethiol. The hybrid CSCNPs can be used as a promising platform for spatiotemporal drug delivery, cell imaging, and theranostics. Based on the same CSCNP platform, flexible functions can be adjusted according to the application needs.

Polymerization-induced proteinosome formation.

In recent years, the fabrication of well-organized proteinosomes has been a popular topic due to the potential applications of the structures in materials science and nanotechnology. A big challenge in the fabrication of proteinosomes is to maintain the structures and the functionalities of proteins on the proteinosomes. In this research, a new concept of polymerization-induced formation of proteinosomes is proposed. In thermal dispersion polymerization of N-isopropyl acrylamide (NIPAM) in the presence of bovine serum albumin (BSA), the growing PNIPAM chains experience phase transition from hydrated coils to dehydrated globules, and the dehydrated PNIPAM chains have hydrophobic interaction with BSA, leading to the formation of hollow proteinosomes. Kinetics studies indicate that there is a transition from the homogeneous polymerization of NIPAM in solution to the heterogeneous polymerization in the proteinosomes. Transmission electron microscopy, atomic force microscopy, confocal laser scanning microscopy and dynamic light scattering all demonstrate the formation of hollow structures. The results of circular dichroism spectroscopy indicate that the secondary structure of BSA remains unchanged in the polymerization process. The formation of proteinosomes is reversible. Upon cooling of the solution to a temperature below the phase transition temperature of PNIPAM, the proteinosomes are dissociated due to the absence of the hydrophobic interaction. The proteinosomes can be used in the encapsulation of hydrophilic compounds in aqueous solution. In this research, not only BSA but also ovalbumin (OVA) is used as a model protein for the fabrication of proteinosomes by the polymerization-induced approach.

Janus Surface Micelles on Silica Particles: Synthesis and Application in Enzyme Immobilization.

In these years, synthesis and applications of Janus structures have aroused great interest for large-scale applications in chemistry and materials science. Up to now, Janus particles with different morphologies and different functionalities have been synthesized in solutions, but the synthesis of Janus particles on solid surfaces has not been touched. In this research, Janus surface micelles (JSMs) are fabricated on the surfaces of silica particles by polymerization induced surface self-assembly (PISSA) approach, and the JSMs are used for enzyme immobilization. Usually, enzyme immobilization should be able to optimize the performance of the immobilized enzymes, and an ideal immobilization system must offer protection to the immobilized enzyme with retained bioactivity. Herein, it is demonstrated that JSMs on silica particles can be used as an ideal platform for the immobilization of enzymes. To prepare JSMs, poly(2-(dimethylamino) ethyl methacrylate) macro chain transfer agent (PDMAEMA-CTA) brushes on silica particles and poly(di(ethylene glycol) methyl ether methacrylate) macro CTA (PDEGMA-CTA) are employed in reversible addition-fragmentation chain transfer dispersion polymerization of styrene. After polymerization, JSMs with polystyrene cores and PDMAEMA/PDEGMA patches on the surfaces are prepared on silica particles. After quaternization reaction, the quaternized PDMAEMA patches are used for the immobilization of enzymes. Experimental results turn out that enhanced bioactivities of the immobilized enzymes are achieved and the enzyme molecules are well protected by surface Janus structures.

Biosurfaces Fabricated by Polymerization-Induced Surface Self-Assembly.

Surface biofunctionalization provides an approach to the fabrication of surfaces with improved biological and clinical performances. Biosurfaces have found increasing applications in many areas such as sensing, cell growth, and disease detection. Efficient synthesis of biosurfaces without damages to the structures and functionalities of biomolecules is a great challenge. Polymerization-induced surface self-assembly (PISSA) provides an effective approach to the synthesis of surface nanostructures with different compositions, morphologies, and properties. In this research, application of PISSA in the fabrication of biosurfaces is investigated. Two different reversible addition-fragmentation chain transfer (RAFT) agents, RAFT chain transfer agent (CTA) on silica particles (SiO2-CTA) and CTA on bovine serum albumin (BSA-CTA), were employed in RAFT dispersion polymerization of N-isopropylacrylamide (NIPAM) in water at a temperature above the lower critical solution temperature (LCST) of poly-(isopropylacrylamide) (PNIPAM). After polymerization, PNIPAM layers with BSA on the top surfaces are fabricated on the surfaces of silica particles. Transmission electron microscopy results show that the average PNIPAM layer thickness increases with monomer conversion. Kinetics study indicates that there is a turn point on a plot of ln([M]0/[M]t) versus polymerization time. After the critical point, surface coassembly of PNIPAM brushes and BSA-PNIPAM bioconjugates is performed on the silica particles. The secondary structure and the activity of BSA immobilized on top of the PNIPAM layers are basically kept unchanged in the PISSA process. To prepare permanently immobilized protein surfaces, PNIPAM layers on silica particles are cross-linked. BSA on the top surfaces presents a reversible "on-off" switching property. At a temperature below the LCST of PNIPAM, the activity of the immobilized BSA is retained; however, the BSA activity decreases significantly at a temperature above the LCST because of the hydrophobic interaction between PNIPAM and BSA. Based on this approach, many different biosurfaces can be fabricated and the materials will find applications in many fields, such as enzyme immobilization, drug delivery, and tissue engineering.

Methionine-Based pH and Oxidation Dual-Responsive Block Copolymer: Synthesis and Fabrication of Protein Nanogels.

In this paper, we synthesized a block copolymer containing pendent thioether functionalities by reversible addition-fragmentation chain transfer polymerization of a tert-butyloxycarbonyl (Boc)-l-methionine-(2-methacryloylethyl)ester (Boc-METMA) monomer using a poly(ethylene glycol) (PEG)-based chain transfer agent. The deprotection of Boc groups resulted in an oxidation and pH dual-responsive cationic block copolymer PEG-b-P(METMA). The block copolymer PEG-b-P(METMA) possessing protonable amine groups was water-soluble at pH < 6.0 and self-assembled to form spherical micelles at pH > 6.0. In the presence of H2O2, the micelles first became highly swollen with time and completely disassembled at last, demonstrating the H2O2-responsive feature because of the oxidation of hydrophobic thioether to hydrophilic sulfoxide. The anticancer drug curcumin (Cur) was entrapped in the polymeric micelles and the Cur-loaded micelles displayed a H2O2-triggered release profile as well as a pH-dependent release behavior, making PEG-b-P(METMA) micelles promising nanocarriers for reactive oxygen species-responsive drug delivery. Taking advantage of the protonated amine groups, the cationic polyelectrolyte PEG-b-P(METMA) formed polyion complex micelles with glucose oxidase (GOx) through electrostatic interactions at pH 5.8. By cross-linking the cores of PIC micelles with glutaraldehyde, the PIC micelles were fixed to generate stable GOx nanogels under physiological conditions. The GOx nanogels were glucose-responsive and exhibited glucose-dependent H2O2-generation activity in vitro and improved storage and thermal stability of GOx. Cur can be encapsulated in the GOx nanogels, and the Cur-loaded GOx nanogels demonstrate the glucose-responsive release profile. The GOx nanogels displayed high cytotoxicity to 4T1 cells and were effectively internalized by the cells. Therefore, these GOx nanogels have potential applications in the areas of cancer starvation and oxidation therapy.

Surface Nanostructures Based on Assemblies of Polymer Brushes.

Studies on the fabrication of hierarchical surface nanostructures have significantly promoted the development of materials with new surface properties and functionalities. In this Minireview, progress on the fabrication of surface nanostructures based on surface organization of polymer brushes are outlined. In the past decades, self-assemblies of polymer brushes, including homopolymer, block-copolymer and mixed-polymer brushes, have aroused great interest in the fields of chemistry and materials science. Recent studies demonstrate that surface co-assemblies of polymer brushes and free polymer chains on solid surfaces are considered as an efficient approach to the preparation of hierarchical nanostructures. Some typical surface nanostructures and properties achieved through surface co-assembly approach are reviewed in this article. Meanwhile, the fundamental problems in the co-assembly approach are discussed and the potential applications of the hierarchical surface nanostructures are presented.

Electrostatic assisted fabrication and dissociation of multi-component proteinosomes.

Self-assembly of proteins into well-organized proteinosomes has attracted great interest due to the potential medical and biological applications of the structures. Herein, a new concept of electrostatic assisted fabrication of proteinosomes is proposed. The self-assembly is performed by using multi-step dialysis approach, where negatively charged bovine serum albumin-poly(N-isopropylacrylamide) (BSA-PNIPAM) bioconjugate and positively charged enzyme (lysozyme or trypsin) are initially dissolved in phosphate buffer (PB) solution at a high salt concentration, and subsequently the protein solution is dialyzed against PB solutions at low salt concentrations, resulting in the formation of biofunctional proteinosomes. Transmission electron microscopy (TEM), cryo-TEM and light scattering results all demonstrate the formation of hollow structures. The wall of a proteinosome is composed of BSA and enzyme (lysozyme or trypsin), and PNIPAM chains of the bioconjugate are in the corona stabilizing the structure. In comparison with the native enzymes, the enzyme molecules in the assemblies basically retain their bioactivities. The proteinosomes formed by BSA-PNIPAM and lysozyme can be dissociated in the presence of trypsin, and those self-assembled by BSA-PNIPAM and trypsin are able to be self-hydrolyzed, resulting in the dissociation of the structures in aqueous solution. The size and morphology changes of the proteinosomes in the hydrolysis are studied.