Chemical Cross-linking Research Articles

An energy-saving structural optimization strategy for high-performance multifunctional graphene films

Graphene has been widely studied for its excellent properties of outstanding conductivity, strength, and flexibility. However, interlayer defects formed during the macroscopic assembly of graphene nanosheets will severely degrade the performance of graphene films. Herein, a strategy combined with trace carboxyl graphene oxide (GO), mild reduction, and chemical crosslinking is employed to minimize the interlayer defects for enhancing both the conductivity and mechanical strength of graphene films. The trace carboxyl feature of GO allows nanosheets to assemble orderly into highly oriented GO films. Subsequently, the mild chemical reduction preserves the highly oriented and packed microstructure. Coupling with further crosslinking by introducing conjugated molecules with pyrene terminal groups (1-pyrene butyric acid-1,6-hexane diamine, PHD) to provide extra π-π bonds, both the mechanical strength and conductivity of graphene films are improved to 548.4 MPa and 1.7 × 105 S m−1, respectively. In addition, the graphene film with only ∼2 μm in thickness possesses an excellent electromagnetic interference shielding effectiveness (EMI SE) of ∼40 dB and a specific shielding effectiveness of up to 74 278 dB cm2 g−1 in a wide frequency range of 8.2–40.0 GHz. Thus, this study provides an energy-saving fabrication route for mechanically strong, effective thermal management and EMI shielding films.

Synthesis of Novel Antimicrobial and Food‐Preserving Hydrogel Nanocomposite Films Based on Carboxymethylcellulose

AbstractThe microbial resistance to the traditional antibiotics causes serious health problems and increases day by day. Thus, there is a pressing need to discover alternative antimicrobial agents of different nature than that of the conventional antibiotics. For this, novel hydrogel nanocomposite films are prepared by chemical crosslinking grafting of carboxymethyl cellulose with N,N′‐methylene‐bis‐acrylamide (CMC‐g‐MBA), which subsequently followed by loading two different weight ratios of titanium oxide nanoparticles (TiO2NPs), such as 3% (CMC‐g‐MBA/TiO2NPs‐3%) and 5% (CMC‐g‐MBA/TiO2NPs‐5%) wt%. Their chemical structure and surface morphology are confirmed using appropriate analytical techniques. Their antimicrobial activity can be arranged as follows: CMC‐g‐MBA/TiO2NPs‐5% composite > CMC‐g‐MBA/TiO2NPs‐3% composite > CMC‐g‐MBA hydrogel. While, the native CMC has no observable antimicrobial effect. CMC‐g‐MBA/TiO2NPs‐3% composite imparts good barrier properties to the coated tomato fruits, delaying their spoilage and protecting them from the environmental microbes compared to the uncoated ones. CMC‐g‐MBA/TiO2NPs‐5% composite is safe on the normal human cells. Thus, incorporation of both MBA and TiO2NPs into CMC greatly develops its antimicrobial activity and provides a wonderful approach to attain promising materials that can efficiently compete with conventional antibiotics.

Assessment of Two Restraint Potentials for Coarse-Grained Chemical-Cross-Link-Assisted Modeling of Protein Structures.

The influence of distance restraints from chemical cross-link mass spectroscopy (XL-MS) on the quality of protein structures modeled with the coarse-grained UNRES force field was assessed by using a protocol based on multiplexed replica exchange molecular dynamics, in which both simulated and experimental cross-link restraints were employed, for 23 small proteins. Six cross-links with upper distance boundaries from 4 Å to 12 Å (azido benzoic acid succinimide (ABAS), triazidotriazine (TATA), succinimidyldiazirine (SDA), disuccinimidyl adipate (DSA), disuccinimidyl glutarate (DSG), and disuccinimidyl suberate (BS3)) and two types of restraining potentials ((i) simple flat-bottom Lorentz-like potentials dependent on side chain distance (all cross-links) and (ii) distance- and orientation-dependent potentials determined based on molecular dynamics simulations of model systems (DSA, DSG, BS3, and SDA)) were considered. The Lorentz-like potentials with properly set parameters were found to produce a greater number of higher-quality models compared to unrestrained simulations than the MD-based potentials, because the latter can force too long distances between side chains. Therefore, the flat-bottom Lorentz-like potentials are recommended to represent cross-link restraints. It was also found that significant improvement of model quality upon the introduction of cross-link restraints is obtained when the sum of differences of indices of cross-linked residues exceeds 150.

Regulation of Cell Adhesion on Physically Crosslinked Hydrogels Composed of Amino Acid‐Based Polymers by Changing Elastic Modulus Using Shape Fix/Memory Properties

AbstractThe elastic modulus of hydrogels is one of the most important factors for controlling cell fate. In this study, hydrogels composed of poly(N‐acryloylglycinamide) (PNAGAm) grafted with arginine (R)–glycine (G)–aspartic acid (D)–serine (S) peptide is designed without a chemical crosslinker. The hydrogels are prepared by the conventional radical copolymerization of N‐acryloylglycinamide with a polymerizable RGDS peptide. The peptide grafting ratio is easily controlled by adjusting the feed composition for polymerization. The hydrogels exhibit thermo‐responsiveness of the upper critical solution temperature type based on the PNAGAm moiety. This characteristic reflects the shape fix/memory properties of hydrogels because of the reversible formation of multiple hydrogen bonds. The cell adhesiveness of the hydrogels drastically improves in the presence of a small amount of the peptide graft. The hydrogels exhibit good biocompatibility; the adhered cells proliferate on the hydrogels, and macrophages do not show activation or inflammation. Surfaces with different elastic modulus regions are successfully constructed on the same hydrogel by using shape fix/memory properties, allowing the regulation of cell adhesion. This hydrogel system offers promising opportunities for the design and application of functional cell scaffolds.

Effects of various cross-linked collagen scaffolds on wound healing in rats model by deep-learning CNN

Scar tissue is connective tissue formed on the wound during the wound-healing process. The most significant distinction between scar tissue and normal tissue is the appearance of covalent cross-linking and the amount of collagen fibers in the tissue. This study investigates the efficacy of four types of collagen scaffolds in promoting wound healing and regeneration in a Sprague-Dawley murine model—the histomorphology analysis of collagen scaffolds and developing a deep learning model for accurate tissue classification. Four female rats (n = 24) groups received collagen scaffolds prepared through physical and chemical crosslinking. Wound healing progress was evaluated by monitoring granulation tissue formation, collagen matrix organization, and collagen fiber deposition, with histological scoring for quantification—the EDC and HA groups demonstrated enhanced tissue regeneration. The EDC and HA groups observed significant differences in wound regeneration outcomes. Deep-learning CNN models with data augmentation techniques were used for image analysis to enhance objectivity. The CNN architecture featured pre-trained VGG16 layers and global average pooling (GAP) layers. Feature visualization using Grad-CAM heatmaps provided insights into the neural network’s focus on specific wound features. The model’s AUC score of 0.982 attests to its precision. In summary, collagen scaffolds can promote wound healing in mice, and the deep learning image analysis method we proposed may be a new method for wound healing assessment.

Ultra-deep reservoirs gel fracturing fluid with stepwise reinforcement network from supramolecular force to chemical crosslinking

Hydraulic fracturing stimulation is a crucial treatment for the development of deep/ultra-deep reservoirs. The success of this treatment is highly dependent on the property of the fracturing fluid, particularly its performance and ability to withstand high pressure without degradation. The high temperature and long wellbore characteristics of deep/ultra-deep reservoirs demand fracturing fluid with high requirements for shear resistance and thermal stability. These attributes are crucial for the effective transportation of proppants over long distances and for minimizing fluid loss. Enhancing the chemical crosslinking intensity can improve the thermal stability of the gel fracturing fluid. However, premature crosslinking can result in frictional pressure loss and mechanical degradation of the fracturing fluid along the long wellbore. In this study, polyacrylamide-based quaternary copolymers with a multi-functional imidazole ring are synthesized. The strong supramolecular force existing between the imidazole ring and carboxylic acid allows for sufficient time intervals before initiating the crosslinking reaction through steric effects, thereby alleviating mechanical degradation and reducing excessive frictional pressure loss. Furthermore, the anti-shear ability of the fracturing fluid is strengthened by a three-dimensional physical network structure. Additionally, thermal/shear stability tests demonstrate that the working temperature of the synthetic polymer (SP) gel fracturing fluid can reach up to 160 °C. Moreover, the fracturing fluid degrades without leaving visible residues under static conditions, which is beneficial to maintaining the integrity of the reservoir for post-fracturing treatment.

Characterization of fish-specific IFNγ-related binding with a unique receptor complex and signaling through a novel pathway.

Unlike mammals, fish express two type II interferons, IFNγ and fish-specific IFNγ (IFNγ-related or IFNγrel). We previously reported the presence of two IFNγrel genes, IFNγrel 1 and IFNγrel 2, which exhibit potent antiviral activity in the Ginbuna crucian carp, Carassius auratus langsdorfii. We also found that IFNγrel 1 increased allograft rejection; however, the IFNγrel 1 receptor(s) and signaling pathways underlying this process have not yet been elucidated. In this study, we examined the unique signaling mechanism of IFNγrel 1 and its receptors. The phosphorylation and transcriptional activation of STAT6 in response to recombinant Ginbuna IFNγrel 1 (rgIFNγrel 1) was observed in Ginbuna-derived cells. Binding of rgIFNγrel 1 to Class II cytokine receptor family members (Crfbs), Crfb5 and Crfb17, which are also known as IFNAR1 and IFNGR1-1, respectively, was detected by flow cytometry. Expression of the IFNγrel 1-inducible antiviral gene, Isg15, was highest in Crfb5- and Crfb17-overexpressing GTS9 cells. Dimerization of Crfb5 and Crfb17 was detected by chemical crosslinking. The results indicate that IFNγrel 1 activates Stat6 through an interaction with unique pairs of receptors, Crfb5 and Crfb17. Indeed, this cascade is distinct from not only that of IFNγ but also that of known IFNs in other vertebrates. IFNs may be classified by their receptor and signal transduction pathways. Taken together, IFNγrel 1 may be classified as a novel type of IFN family member in vertebrates. Our findings provide important information on interferon gene evolution in bony fish.

A microscale 3D organ on a chip for recapitulating reciprocal neuroendocrine crosstalk between the hypothalamus and the pituitary gland

Conventional 2D or even recently developed 3D in vitro culture models for hypothalamus and pituitary gland cannot successfully recapitulate reciprocal neuroendocrine communications between these two pivotal neuroendocrine tissues known to play an essential role in controlling the body’s endocrine system, survival, and reproduction. In addition, most current vitro culture models for neuroendocrine tissues fail to properly reflect their complex multicellular structure. In this context, we developed a novel microscale chip platform, termed the ‘hypothalamic–pituitary (HP) axis-on-a-chip,’ which integrates various cellular components of the hypothalamus and pituitary gland with biomaterials such as collagen and hyaluronic acid. We used non-toxic blood coagulation factors (fibrinogen and thrombin) as natural cross-linking agents to increase the mechanical strength of biomaterials without showing residual toxicity to overcome drawbacks of conventional chemical cross-linking agents. Furthermore, we identified and verified SERPINB2 as a reliable neuroendocrine toxic marker, with its expression significantly increased in both hypothalamus and pituitary gland cells following exposure to various types of toxins. Next, we introduced SERPINB2-fluorescence reporter system into loaded hypothalamic cells and pituitary gland cells within each chamber of the HP axis on a chip, respectively. By incorporating this SERPINB2 detection system into the loaded hypothalamic and pituitary gland cells within our chip platform, Our HP axis-on-chip platform can better mimic reciprocal neuroendocrine crosstalk between the hypothalamus and the pituitary gland in the brain microenvironments with improved efficiency in evaluating neuroendocrine toxicities of certain drug candidates.

A Tent-Inspired Portable Solar-Driven Water Purification Device for Wilderness Explorers.

Wilderness adventure favored by many enthusiasts often endanger lives due to lacking freshwater or drinking contaminated water. Therefore, compared to the inefficient methods of filtration, steaming, and direct solar heating, it is of great meaningfulness to develop a solar-driven water purification device with efficiency, lightweight, portability, and multi-water-quality purification by taking full advantage of solar-driven interfacial evaporation. Here, a tent-inspired portable solar-driven water purification device consisting of Janus-structured bacterial cellulose aerogel (JBCA) solar evaporator and tent-type condensation recovery device is reported. For the JBCA solar evaporator, it is prepared from biomass bacterial cellulose (BC) as raw material and hydroxylated carbon nanotubes (HCNT) as photothermal material, and the Janus property is achieved by the assistance of hydrophobic and hydrophilic chemical cross-linking. It exhibits lightweight, unibody, high photothermal conversion, efficient evaporation, and multi-water-quality purification capability for representative seawater, urine, and bacterial river water. For the tent-type condensation recovery device, it is based on the prototype of tent and uses flexible ultra-transparent polyvinyl chloride (PVC) film as raw material. Thanks to the rational prototype and material selection, it displays outstanding portability and lightweight through the folding/unfolding method. Therefore, the designed tent-inspired portable solar-driven water purification device demonstrates great potential application in wilderness exploration.

Universal Strategy with Structural and Chemical Crosslinking Interface for Efficient and Stable Perovskite Solar Cells

AbstractDue to the limited interface contact and weak interfacial interaction, planar heterojunction perovskite solar cells (PSCs) have space for further improvement. Herein, a structural and chemical crosslinking interface is proposed and constructed by introducing an extra layer, which blends tin dioxide (SnO2) nanoparticles with chloride salts. Since the incorporated materials can be dissolved during the fabrication of perovskite, the quality of perovskite films is improved, leading to larger grain size and reduced trap‐state density. Also, more chloride ions at the SnO2/perovskite interface are observed and the interaction between Cl− and Sn4+ is confirmed. It results in more pronounced n‐type SnO2 with better conductivity and deeper conduction bands, leading to preferable energy level alignment between SnO2 and perovskite. Consequently, the open‐circuit voltage and fill factor of the devices increase, and target cells present better stability, retaining 98% of initial efficiencies after >10 000 h storage in dry air (≈5% relative humidity) and maintaining 85.50% of the initial efficiency after 1000 h of operation under light. This strategy enables the achievement of 25.28% efficiency with a low bandgap (1.53 eV) perovskite composition, and it is confirmed to be universal when other related materials are utilized.

Tri-network, physical cross-linked sodium alginate hydrogel with strongly self-healing capacity and highly tough, for wearable flexible devices

Physical cross-linked hydrogels offer advantageous self-healing and easy preparation for broad application in wearable flexible devices. However, compared to chemical crosslinking, physical crosslinking is relatively fragile, yielding weak mechanical properties that hinder further development. To solve this problem, we polymerized [2-(methacryloyloxy)ethyl]dimethyl-(propyl3-sulfonate) ammonium hydroxide (DMAPS) into SA-PMAA hydrogels to form a three-network physical cross-linked hydrogel with high strength and superior self-healing ability. PDMAPS possesses dipolar-dipole action between lateral base units and self-associating ability. By introducing PDMAPS into the SA-PMAA double network structure, we created a hydrogel with excellent self-healing properties and mechanical strength. The hydrogel boasts a mechanical strength of 41 KPa, a maximum elongation at break of 820 %, and 94.7 % self-healing efficiency after 12 h, surpassing most hydrogels. With its polar groups, the hydrogel offers great compatibility with most aqueous slurries containing inorganic fillers, making it a material with immense potential as a polyelectrolyte and broad applications in the field of biosensing and wearable flexible devices.

Physically cross-linked scaffold composed of hydroxyapatite-chitosan-alginate-polyamide has potential to trigger bone regeneration in craniofacial defect

Abstract Recently, the fabrication of a scaffold from biomaterials has been increased due to the lack of adequate natural bone for grafting. In this study, hydroxyapatite-chitosan-alginate-polyamide (HCAP) synthetic scaffold was fabricated using the thermally induced phase separation (TIPS) technique. The scaffold was cross-linked with either a chemical cross-linker (calcium chloride, 2-hydroxyethyl methacrylate (HEMA), or glutaraldehyde (GTA)) or a physical cross-linker (gamma irradiation (IR)) resulting in scaffolds HCAP-CaCl2, HCAP-HEMA, HCAP-GTA, and HCAP-IR, respectively. The cross-linked scaffolds were characterized based on physicochemical properties, cytotoxicity, and biocompatibility. HCAP-GTA showed the highest density and the lowest swelling ratio and biodegradation rate closely matching with those of the HCAP-IR. Porosity and density of the HCAP, HCAP-IR, and HCAP-GTA scaffolds were 92.14, 87.26, and 83.33 %, and 0.241, 0.307, and 0.335 g/cm3, respectively. The swelling ratio for the same scaffolds was 149, 110, and 108 % after 72 h of observation. Brine shrimp cytotoxicity and RBC biocompatibility assay confirmed the non-toxic nature of HCAP-IR and HCAP-GTA scaffolds. The HCAP-IR scaffold was tested for bone regeneration in the rabbit mandible defect model. Histological analysis revealed the regeneration of new bone and restoration of bone defect at the site of injury. These findings indicate that radiation induced physically cross-linked HCAP scaffold could be used as an alternative in bone defect replacement therapy.

A Sustainable Dual Cross-Linked Cellulose Hydrogel Electrolyte for High-Performance Zinc-Metal Batteries

HighlightsA sustainable dual cross-linked cellulose hydrogel with excellent mechanical strength was fabricated from aqueous alkali hydroxide/urea solution using a sequential chemical and physical cross-linking strategy.The hydrogel electrolyte effectively suppresses dendrites growth and side reactions to achieve a stable Zn anode (over 2000 h for Zn||Zn cell), which are proved by a multi-perspective and in-depth mechanism investigation.The hydrogel electrolyte is easily accessible and biodegradable, making the zinc batteries attractive in terms of scalability and sustainability.

Structural mechanisms of autoinhibition and substrate recognition by the ubiquitin ligase HACE1.

Ubiquitin ligases (E3s) are pivotal specificity determinants in the ubiquitin system by selecting substrates and decorating them with distinct ubiquitin signals. However, structure determination of the underlying, specific E3-substrate complexes has proven challenging owing to their transient nature. In particular, it is incompletely understood how members of the catalytic cysteine-driven class of HECT-type ligases (HECTs) position substrate proteins for modification. Here, we report a cryogenic electron microscopy (cryo-EM) structure of the full-length human HECT HACE1, along with solution-based conformational analyses by small-angle X-ray scattering and hydrogen-deuterium exchange mass spectrometry. Structure-based functional analyses in vitro and in cells reveal that the activity of HACE1 is stringently regulated by dimerization-induced autoinhibition. The inhibition occurs at the first step of the catalytic cycle and is thus substrate-independent. We use mechanism-based chemical crosslinking to reconstitute a complex of activated, monomeric HACE1 with its major substrate, RAC1, determine its structure by cryo-EM and validate the binding mode by solution-based analyses. Our findings explain how HACE1 achieves selectivity in ubiquitinating the active, GTP-loaded state of RAC1 and establish a framework for interpreting mutational alterations of the HACE1-RAC1 interplay in disease. More broadly, this work illuminates central unexplored aspects in the architecture, conformational dynamics, regulation and specificity of full-length HECTs.

Mapping Protein-Protein Interactions at Birth: Single-Particle Cryo-EM Analysis of a Ribosome-Nascent Globin Complex.

Interactions between ribosome-bound nascent chains (RNCs) and ribosomal components are critical to elucidate the mechanism of cotranslational protein folding. Nascent protein-ribosome contacts within the ribosomal exit tunnel were previously assessed mostly in the presence of C-terminal stalling sequences, yet little is known about contacts taking place in the absence of these strongly interacting motifs. Further, there is nearly no information about ribosomal proteins (r-proteins) interacting with nascent chains within the outer surface of the ribosome. Here, we combine chemical cross-linking, single-particle cryo-EM, and fluorescence anisotropy decays to determine the structural features of ribosome-bound apomyoglobin (apoMb). Within the ribosomal exit tunnel core, interactions are similar to those identified in previous reports. However, once the RNC enters the tunnel vestibule, it becomes more dynamic and interacts with ribosomal RNA (rRNA) and the L23 r-protein. Remarkably, on the outer surface of the ribosome, RNCs interact mainly with a highly conserved nonpolar patch of the L23 r-protein. RNCs also comprise a compact and dynamic N-terminal region lacking contact with the ribosome. In all, apoMb traverses the ribosome and interacts with it via its C-terminal region, while N-terminal residues sample conformational space and form a compact subdomain before the entire nascent protein sequence departs from the ribosome.

Cross-Linking Methods of the Silk Protein Hydrogel in Oral and Craniomaxillofacial Tissue Regeneration.

Craniomaxillofacial tissue defects are clinical defects involving craniomaxillofacial and oral soft and hard tissues. They are characterized by defect-shaped irregularities, bacterial and inflammatory environments, and the need for functional recovery. Conventional clinical treatments are currently unable to achieve regeneration of high-quality oral craniomaxillofacial tissue. As a natural biomaterial, silk fibroin (SF) has been widely studied in biomedicine and has broad prospects for use in tissue regeneration. Hydrogels made of SF showed excellent water retention, biocompatibility, safety and the ability to combine with other materials. To gain an in-depth understanding of the current development of SF, this article reviews the structure, preparation and application prospects in oral and craniomaxillofacial tissue regenerative medicine. It first briefly introduces the structure of SF and then summarizes the principles, advantages and disadvantages of the different cross-linking methods (physical cross-linking, chemical cross-linking and double network structure) of SF. Finally, the existing research on the use of SF in tissue engineering and the prospects of using SF with different cross-linking methods in oral and craniomaxillofacial tissue regeneration are also discussed. This review is intended to show the advantages of SF hydrogels in tissue engineering and provides theoretical support for establishing novel and viable silk protein hydrogels for regeneration.

Tuning the Mechanical and Functional Properties of Pea Protein-Based Bioplastics via Different Physical and Chemical Cross-Linking Methods

Tuning the Mechanical and Functional Properties of Pea Protein-Based Bioplastics via Different Physical and Chemical Cross-Linking Methods

Construction of a Soft Antifouling PAA/PSBMA Hydrogel Coating with High Toughness and Low Swelling through the Dynamic Coordination Bonding Provided by Al(OH)3 Nanoparticles.

Marine biofouling, resulting from the adhesion of marine organisms to ship surfaces, has long been a significant issue in the maritime industry. In this paper, we focused on utilizing soft and hydrophilic hydrogels as a potential approach for antifouling (AF) coatings. Acrylic acid (AA) with a polyelectrolyte effect and N-(3-sulfopropyl)-N-(methacryloxyethyl)-N,N-dimethylammonium betaine (SBMA) with an antipolyelectrolyte effect were selected as monomers. By adjusting the monomer ratio, we were able to create hydrogel coatings that exhibited low swelling ratio in both fresh water and seawater. The Al(OH)3 nanoparticle, as a physical cross-linker, provided better mechanical properties (higher tensile strength and larger elongation at break) than the chemical cross-linker through the dynamic coordination bonds and plentiful hydrogen bonds. Additionally, we incorporated trehalose into the hydrogel, enabling the repair of the hydrogel network through covalent-like hydrogen bonding. The zwitterion compound SBMA endowed the hydrogel with excellent AF performance. It was found that the highest SBMA content did not lead to the best antibacterial performance, as bacterial adhesion quantity was also influenced by the charge of the hydrogel. The hydrogel with appropriate SBMA content being close to electrical neutrality exhibits the strongest zwitterionic property of PSBMA chains, resulting in the best antibacterial adhesion performance. Furthermore, the pronounced hydrophilicity of SBMA enhanced the lubrication of the hydrogel surface, thereby reducing the friction resistance when applied to the hull surface during ship navigation.

Multifunctional sodium lignosulfonate/xanthan gum/sodium alginate/polyacrylamide ionic hydrogels composite as a high-performance wearable strain sensor

Recently, conductive hydrogels have shown great promise in flexible electronics and are ideal materials for the preparation of wearable strain sensors. However, developing a simple method to produce conductive hydrogels with excellent mechanical properties, self-adhesion, transparency, anti-freezing, and UV resistance remains a significant challenge. A novel sodium lignosulfonate/xanthan gum/sodium alginate/polyacrylamide/Zn2+/DMSO (SLS/XG/SA/PAM/Zn2+/DMSO) ionic conductive hydrogel was developed using a one-pot method. The resulting ionic conductive hydrogels have excellent mechanical properties (stress: 0.13 MPa, strain: 1629 %), high anti-fatigue properties, self-adhesion properties (iron: 7.37 kPa, pigskin: 4.74 kPa), anti-freezing (freezing point: −33.49 °C) and UV resistance by constructing a chemical and physical hybrid cross-linking network. In particular, the conductivity of G hydrogel reached 6.02 S/m at room temperature and 5.52 S/m at −20 °C. Thus, the hydrogel was assembled into a flexible sensor that could distinguish a variety of large and small scales human movements, such as joint bending, swallowing and speaking in real time with high stability and sensitivity. Moreover, the hydrogel could be used as electronic skin just like human skin and touch screen pen to write.

Rapid Profiling of the Glycosylation Effects on the Binding of SARS-CoV-2 Spike Protein to Angiotensin-Converting Enzyme 2 Using MALDI-MS with High Mass Detection.

The spike protein receptor-binding domain (RBD) of SARS-CoV-2 binds directly to angiotensin-converting enzyme 2 (ACE2), mediating the host cell entry of SARS-CoV-2. Both spike protein and ACE2 are highly glycosylated, which can regulate the binding. Here, we utilized high-mass MALDI-MS with chemical cross-linking for profiling the glycosylation effects on the binding between RBD and ACE2. Overall, it was found that ACE2 glycosylation affects the binding more strongly than does RBD glycosylation. The binding affinity was improved after desialylation or partial deglycosylation (N690) of ACE2, while it decreased after degalactosylation. ACE2 can form dimers in solution, which bind more tightly to the RBD than the ACE2 monomers. The ACE2 dimerization and the binding of RBD to dimeric ACE2 can also be improved by the desialylation or deglycosylation of ACE2. Partial deglycosylation of ACE2 increased the dimerization of ACE2 and the binding affinity of RBD and ACE2 by more than a factor of 2, suggesting its high potential for neutralizing SARS-CoV-2. The method described in the work provided a simple way to analyze the protein-protein interaction without sample purification. It can be widely used for rapid profiling of glycosylation effects on protein-protein interaction for glycosylation-related diseases and the study of multiple interactions between protein and protein aggregates in a single system.