Encapsulation of multiple enzymes within a microgel via water-in-water emulsions for enzymatic cascade reactions.

Enzyme-loaded spherical microgels with diameters of several micrometers have been explored for use in therapeutic microreactors and biosensors. Conventional preparation strategies for enzyme-loaded microgels utilized water-in-oil emulsions or flow chemistry techniques. The former damage enzyme activity using organic solvents and the latter are expensive and difficult to expand because of the complex system. In this study, we present a simple strategy for creating multiple enzyme-loaded gelatin-based microgels with tunable diameters in a single flask. This strategy was based on our finding that enzymes spontaneously partitioned in a dispersed methacryloyl gelatin aqueous solution in a poly(vinylpyrrolidone) (WGelMA/WPVP) aqueous solution. The method achieved an encapsulation efficiency of over 70% even with four types of enzymes and retained their activity owing to the full aqueous system. Additionally, the encapsulated ß-galactosidase activity was maintained for 24 hours at pH 6, although naked ß-galactosidase lost approximately 60% of its activity, which was superior to that of previous enzyme-loaded gelatin gels. Moreover, this simple method enabled the production of 10 g-scale or more microgels in one batch. We also demonstrated that multiple enzyme-loaded gelatin microgels functioned as cascade microreactors for lactose and glucose sensing. This versatile strategy enables the production of enzyme-loaded microgels while maintaining the enzyme activity using very low technologies. This result contributes to the easy preparation of enzyme-loaded microgels and their applications in the biomedical and green catalytic fields.

Well-Defined Anisotropic Self-Assembly from Peptoids and Their Biomedical Applications.

Peptoids, or poly(N-substituted glycine)s, hold great promise in biomedical applications because of their biocompatibility, precise synthesis via conventional peptide-mimicking methods, and readily tunable side chains, which facilitate the control of hydrophobicity and crystallinity. In the past decade, peptoids have been used to create well-defined self-assemblies such as vesicles, micelles, sheets, and tubes, which have been scrutinized at the atomic scale using cutting-edge analytical techniques. This review highlights recent advancements in peptoid synthesis strategies and the development of noteworthy one- or two-dimensional anisotropic self-assemblies, i. e., nanotubes and nanosheets, exhibiting well-ordered molecular arrangements. These anisotropic self-assemblies are formed through the crystallization of peptoid side chains, which can be effortlessly modified via simple synthesis approaches. Moreover, leveraging the protease resistance of peptoids, various biomedical applications are discussed (including phototherapy, enzymatic mimetics, bio-imaging, and biosensing) that capitalize on the unique properties of anisotropic self-assembly.

Glycopeptoid nanospheres: glycosylation-induced coacervation of poly(sarcosine).

Conjugation of maltopentaose to water-soluble homo-poly(sarcosine) induced self-association and formed nanospheres (-150 nm) in water although homo-poly(sarcosine) was water-soluble and did not form any aggregates. Fluorescent probe experiments showed that the spheres were non-ionic glycopeptoid coacervate-like particles with both hydrophobic and hydrophilic domains inside.

Thermoresponsive Carbohydrate-b-Polypeptoid Polymer Vesicles with Selective Solute Permeability and Permeable Factors for Solutes.

Solute-permeable polymer vesicles are structural compartments for nanoreactors/nanofactories in the context of drug delivery and artificial cells. We previously proposed design guidelines for polymers that form solute-permeable vesicles, yet we did not provide enough experimental verification. In addition, the fact that there is no clear factor for identifying permeable solutes necessitates extensive trial and error. Herein, we report solute-permeable polymer vesicles based on an amphiphilic copolymer, thermoresponsive oligosaccharide-block-poly(N-n-propylglycine). The introduction of a thermoresponsive polymer as a hydrophobic segment into amphiphilic polymers is a viable approach to construct solute-permeable polymer vesicles. We also demonstrate that the polymer vesicles are preferentially permeable to cationic and neutral fluorophores and are hardly permeable to anionic fluorophores due to the electrostatic repulsion between the bilayer and anionic fluorophores. In addition, the permeability of neutral fluorophores increases with the increasing log P value of the fluorophores. Thus, the electrical charge and log P value are important factors for membrane permeability. These findings will help researchers develop advanced nanoreactors based on permeable vesicles for a broad range of fundamental and biomedical applications.

A Novel Surface Modification and Immobilization Method of Anti-CD25 Antibody on Nonwoven Fabric Filter Removing Regulatory T Cells Selectively.

Anti-CD25 antibodies were immobilized on polypropylene (PP) nonwoven fabrics to specifically remove mouse regulatory T cells (Tregs) from mouse spleen cells. PP fibers were coated with peptide nanosheets, which were prepared by self-assembling of a mixture of X-poly(sarcosine)-b-(l-Leu-Aib)6 (X: glycolic acid or a phenylboronic acid) and Y-poly(sarcosine)-b-(d-Leu-Aib)6 (Y: glycolic acid or diazirine derivative). Anti-CD25 antibodies were immobilized by covalent linking between the sugar moiety of the antibody and the phenylboronic acid group on the peptide nanosheet. The removal rate of mouse Tregs from the mouse spleen cells was more than 95% only by passing the filters, while the nonspecific removal rates of other cells were less than 15%. The coating of peptide nanosheets on PP fibers was very effective to provide a suitable environment for the immobilized antibody to interact with the counterpart cells while the coating suppressed nonspecific adsorption of other cells.