Platform For Immobilization Research Articles

Filamentous fungal pellets as versatile platforms for cell immobilization: developments to date and future perspectives

Filamentous fungi are well-known for their efficiency in producing valuable molecules of industrial significance, but applications of fungal biomass remain relatively less explored despite its abundant and diverse opportunities in biotechnology. One promising application of mycelial biomass is as a platform to immobilize different cell types such as animal, plant, and microbial cells. Filamentous fungal biomass with little to no treatment is a sustainable biomaterial which can also be food safe compared to other immobilization supports which may otherwise be synthetic or heavily processed. Because of these features, the fungal-cell combination can be tailored towards the targeted application and be applied in a variety of fields from bioremediation to biomedicine. Optimization efforts to improve cell loading on the mycelium has led to advancements both in the applied and basic sciences to understand the inter- and intra-kingdom interactions. This comprehensive review compiles for the first time the current state of the art of the immobilization of animal, yeast, microalgae, bacteria, and plant cells in filamentous fungal supports and presents outlook of applications in intensified fermentations, food and biofuel production, and wastewater treatment. Opportunities for further research and development were identified to include elucidation of the physical, chemical, and biological bases of the immobilization mechanisms and co-culture dynamics; expansion of the cell-fungus combinations investigated; exploration of previously unconsidered applications; and demonstration of scaled-up operations. It is concluded that the potential exists to leverage the unique qualities of filamentous fungus as a cellular support in the creation of novel materials and products in support of the circular bioeconomy.

Multi-walled carbon nanotubes functionalized with a new Schiff base containing phenylboronic acid residues: applicationtothe development of a bienzymatic glucose biosensor using a response surface methodology approach.

Aninnovative supramolecular architectureis reportedfor bienzymatic glucose biosensing based on the use of a nanohybrid made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with a Schiff base modified with two phenylboronic acid residues (SB-dBA) as platform for the site-specific immobilization of the glycoproteins glucose oxidase (GOx) and horseradish peroxidase (HRP). The analytical signal was obtained from amperometric experiments at - 0.050V in the presence of 5.0 × 10-4M hydroquinone as redox mediator. The concentration of GOx and HRP and the interaction time between the enzymes and the nanohybrid MWCNT-SB-dBA deposited at glassy carbon electrodes (GCEs) were optimized through a central composite design (CCD)/response surface methodology (RSM). The optimal concentrations of GOx and HRP were 3.0mgmL-1 and 1.50mgmL-1, respectively, while the optimum interaction time was 3.0min. The bienzymatic biosensor presented a sensitivity of (24 ± 2) × 102 µA dL mg-1 ((44 ± 4) × 102 µA M-1), a linear range between 0.06mg dL-1 and 21.6mg dL-1 (3.1µM-1.2mM) (R2 = 0.9991),and detection and quantification limits of 0.02mg dL-1 (1.0µM) and 0.06mg dL-1 (3.1µM), respectively. The reproducibility for five sensors prepared with the same MWCNT-SB-dBA nanohybrid was 6.3%, while the reproducibility for sensors prepared with five different nanohybrids and five electrodes each was 7.9%. The GCE/MWCNT-SB-dBA/GOx-HRP was successfully used for the quantification of glucose in artificial human urine and commercial human serum samples.

An electrochemical biosensor for sensitive detection of live Salmonella in food via MXene amplified methylene blue signals and electrostatic immobilization of bacteriophages.

Anovel bacteriophage-targeted electrochemical biosensor designed for accurate and quantitative detection of live Salmonella in food samplesis presented. The biosensor is simply constructed by electrostatic immobilizing bacteriophages on MXene-nanostructured electrodes. MXene, renowned for its high surface area, biocompatibility, and conductivity, serves as an ideal platform for bacteriophage immobilization. This allows for a high-density immobilization of bacteriophage particles, achieving approximately 71 pcs μm-2. Remarkably, the bacteriophages immobilized MXene nanostructured electrodes still maintain their viability and functionality, ensuring their effectiveness in pathogen detection. Therefore, the proposed biosensor exhibited enhanced sensitivity with a low limit of detection (LOD) of 5CFUmL-1. Notably, the biosensor shows excellent specificity in the presence of other bacteria that commonly contaminate food and can distinguish live Salmonella from a mixed population. Furthermore, it is applicable in detecting live Salmonella in food samples, which highlights its potential in food safety monitoring. This biosensor offers simplicity, convenience, and suitability for resource-limited environments, making it a promising tool for on-site monitoring of foodborne pathogenic bacteria.

Laccase Immobilized on Arginine-Functionalized Boron Nitride Nanosheets for Enhanced Atrazine Degradation.

Enzyme-mediated systems have been widely employed for the biotransformation of environmental contaminants. However, the catalytic performance of free enzymes is restricted by the rapid loss of their catalytic activity, stability, and reusability. In this work, we developed an enzyme immobilization platform by elaborately anchoring fungal laccase onto arginine-functionalized boron nitride nanosheets (BNNS-Arg@Lac). BNNS-Arg@Lac showcased ∼75% immobilization yield and enhanced stability against fluctuating pH values and temperatures, along with remarkable reusability across six consecutive cycles, outperforming free natural laccase (nlaccase). A model pollutant, atrazine, was selected for a proof-of-concept demonstration, given the substantial environmental and public health concerns in agriculture runoff. BNNS-Arg@Lac-catalyzed atrazine degradation rate was nearly twice that of nlaccase. Moreover, BNNS-Arg@Lac consistently demonstrated superior atrazine degradation in synthetic agricultural wastewater and various mediator systems compared to nlaccase. Comprehensive product analysis unraveled distinct degradation pathways for BNNS-Arg@Lac and nlaccase. Overall, this research provides a foundation for the future development of enzyme-nanomaterial hybrids for degrading environmental chemicals and may unlock new potential for green and efficient resource recovery and waste management strategies.

Copper-loaded ZIF-8-based immunosensor for early diagnosis of chronic kidney disease by detecting neutrophil gelatinase-associated lipocalin (NGAL) biomarker

Metal-organic frameworks (MOFs) is a promising contender among the nanomaterials for biosensors due to their special properties, including high surface area, tunable porosity, and adaptable functionalization competences. Zeolitic imidazolate frameworks (ZIFs), a subclass of MOFs, have drawn precise attention for their excellent chemical stability and ease of synthesis. This study, for the first time, concentrates on the preparation and characterization of a Cu-loaded ZIF-8-based immunosensor for the detection of neutrophil gelatinase-associated lipocalin (NGAL), a crucial biomarker for chronic kidney disease (CKD). Loading copper (Cu) into the ZIF-8 framework improves its electrochemical properties, conductivity, and surface area, leading it an ideal platform for antibody immobilization and NGAL detection. The immunosensor utilized Staphylococcal Protein A (SPA) IgG binding protein for antibody immobilization, providing an alternative to traditional crosslinking chemistry, enhancing biosensor functionality. Structural analysis, morphological assessment, elemental mapping, and stoichiometric analysis confirmed the successful synthesis of Cu-loaded ZIF-8 nanocomposite. Assessment of the electrochemical performance of the immunosensor demonstrated sufficiently low limit of detection (80 pg mL−1) over a wide range of 0.1 ng mL−1 to 1000 ng mL−1 of NGAL along with superior selectivity, reproducibility, stability, and clinical feasibility. This advancement contributes to the rising knowledge on MOF-based biosensors and holds promise for future applications in disease diagnosis and healthcare monitoring.

One-pot Functionalization for the Preparation of Cobaltocene-Modified Redox-Responsive Porous Microparticles.

Porous organic cobaltocenium-containing particles are scarce in literature but highly interesting for their electrochemical properties and reusability in, for example, catalysis or magnetic systems. In this work, we present a scalable one-pot strategy to introduce tailorable amounts of cobaltocenium on a porous substrate, adjusting the electrochemical switching capability. For this purpose, 3-(triethoxysilyl)propan-1-amine (APTES) and ethynyl cobaltocenium hexafluorophosphate is used as functionalization agents for in-situ catalyst-free hydroamination, followed by silane condensation at the particles' surface. Functionalized particles are characterized by attenuated total reflection infrared spectroscopy (ATR-IR), thermogravimetric analysis (TGA), laser scanning confocal microscopy (LSCM), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), inductively coupled plasma mass spectrometry (ICP-MS), powder X-ray diffraction (PXRD) and cyclic voltammetry (CV) showing excellent control over the degree of functionalization, i. e., the added cobaltocenium reagents. The electrochemical stability and good addressability while preserving the porous structure are shown. By utilizing higher amounts of APTES, the overall cobaltocenium amount can be reduced in favor of additional amine groups, strongly affecting the electrochemical behavior, making this functionalization strategy a good platform for metallopolymer immobilization and tailored functionalization. Additionally, thermal treatment of the synthesized metallopolymer microparticles paves the way to magnetic properties with tailorable microporous architectures for end-of-life and upcycling aspects.

Enhancing cellulose hydrolysis via cellulase immobilization on zeolitic imidazolate frameworks using physical adsorption.

This study investigates the immobilization of cellulase on zeolitic imidazolate frameworks (ZIFs) by physical adsorption, specifically the ZIF-8-NH2 and Fe3O4@ZIF-8-NH2, to enhance enzymatic hydrolysis efficiency. The immobilization process was thoroughly analyzed, including optimization of conditions and characterization of ZIF carriers and immobilized enzymes. The impacts on the catalytic activity of cellulase under various temperatures, pH levels, and storage conditions were examined. Additionally, the reusability of the immobilized enzyme was assessed. Results showed the cellulase immobilized on Fe3O4@ZIF-8-NH2 exhibited a high loading capacity of 339.64 mg/g, surpassing previous studies. Its relative enzymatic activity was found to be 71.39%. Additionally, this immobilized enzyme system demonstrates robust reusability, retaining 68.42% of its initial activity even after 10 cycles. These findings underscore the potential of Fe3O4@ZIF-8-NH2 as a highly efficient platform for cellulase immobilization, with promising implications for lignocellulosic biorefinery.

Electrochemical cDNA-sensing of miRNA-137 for detection of Alzheimer's disease using CuO and PEI deposited Au-Pd bimetallic nanoparticles

Alzheimer's disease (AD) is the third most predominantly occurring disease worldwide that is characterized by progressive deterioration of brain cells and change in behaviour, personality, orientation of time and space, leading to functional disability which affects the daily life of patient. We report an electrochemical complementary DNA (cDNA)-sensor for detection of AD by using blood biomarker, micro-ribonucleic acid-137 (miRNA-137). In this context, the gold-palladium/polyethyleneimine@copperoxide nanocomposite-coated fluorine tin oxide (Au-Pd/PEI@CuO/FTO) electrode is used as an immobilization platform for complementary DNA (cDNA) against the target miRNA-137. The thiolated-cDNA (Thi-cDNA) and Au-Pd bimetallic particles interact through the stable electrostatic binding. The fabricated Au-Pd/PEI@CuO/FTO was characterized by field emission scanning electron microscopy, cyclic voltammetry, and electrochemical Impendence Spectroscopy. The sensor showed response time of 20 min with the linear range of 1fM to 100 nM and limit of detection as 0.164pM. The recovery of the cDNA-sensor for spiked human serum sample ranged from 99.3 % to 103.28 % with RSD value of 0.62 % to 2.81 %. The easy-to-use procedure, cost effectiveness, compactness, long shelf life (28 days) and high selectivity of cDNA-sensor towards miRNA-137 over even the one-base pair mismatch recommends the sensor's portability and real-life usability for AD detection.

Patterned graphene: An effective platform for adsorption, immobilization, and destruction of SARS-CoV-2 Mpro

To address the ongoing challenges posed by the SARS-CoV-2 and potentially stronger viruses in the future, the development of effective methods to fabricate patterned graphene (PG) and other precisely functional products has become a new research frontier. Herein, we modeled the “checkerboard” graphene (CG) and stripped graphene (SG) as representatives of PG, and studied their interaction mechanism with the target protein (Mpro) by molecular dynamics simulation. The calculation results on the binding strength and the root mean square deviation values of the active pocket revealed that PG is an effective platform for adsorption, immobilization, and destruction of Mpro. Specifically, CG is found to promote disruption of the active pocket for Mpro, but the presence of “checkerboard” oxidized regions inhibits the adsorption of Mpro. Meanwhile, the SG can effectively confine Mpro within the non-oxidized strips and enhances their binding strength, but doesn’t play well on disrupting the active pocket. Our work not only elucidates the biological effects of PGs, but also provides guidance for their targeted and precise utilization in combating the SARS-CoV-2.

Designable immobilization of D-allulose 3-epimerase on bimetallic organic frameworks based on metal ion compatibility for enhanced D-allulose production

D-allulose, a low-calorie rare sugar catalyzed by D-allulose 3-epimerase (DAE), is highly sought after for its potential health benefits. However, poor reusability and stability of DAE limited its popularization in industrial applications. Although metal-organic frameworks (MOFs) offer a promising enzyme platform for enzyme immobilization, developing customized strategies for MOF immobilization of enzymes remains challenging. In this study, we introduce a designable strategy involving the construction of bimetal-organic frameworks (ZnCo-MOF) based on metal ions compatibility. The DAE@MOFs materials were prepared and characterized, and the immobilization of DAE and the enzymatic characteristics of the MOF-immobilized DAE were subsequently evaluated. Remarkably, DAE@ZnCo-MOF exhibited superior recyclability which could maintain 95 % relative activity after 8 consecutive cycles. The storage stability is significantly improved compared to the free form, with a relative activity of 116 % remaining after 30 days. Molecular docking was also employed to investigate the interaction between DAE and the components of MOFs synthesis. The results demonstrate that the DAE@ZnCo-MOF exhibited enhanced catalytic efficiency and increased stability. This study introduces a viable and adaptable MOF-based immobilization strategy for enzymes, which holds the potential to expand the implementation of enzyme biocatalysts in a multitude of disciplines.

L-cysteine-coated magnetite nanoparticles as a platform for enzymes immobilization: Amplifying biocatalytic activity of Candida antarctica Lipase A

This study presents the synthesis of magnetite nanoparticles coated with L-cysteine (Fe3O4@LC) and their subsequent utilization as a support matrix for the immobilization of Candida antarctica Lipase A (CALA). The immobilization process involved physical interactions and covalent bonding mediated by glutaraldehyde. Comprehensive characterization was conducted using techniques including X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), and vibrating sample magnetometry (VSM), providing compelling evidence of a successful synthesis. Under optimized conditions, employing 1 mg of protein per gram of support, pH 7 in a 25 mM phosphate buffer, and a 5 h reaction at 25 °C, immobilization on glutaraldehyde-activated support achieved an impressive yield of 85.0 % ± 2.6, accompanied by a specific activity of 212.5 ± 1.3 U/g, outperforming the physical adsorption approach. Remarkably, the immobilized enzyme exhibited higher activity than the free enzyme at alkaline and acidic pH levels. Furthermore, thermal and pH inactivation studies revealed that the biocatalyst's half-life exceeded that of free CALA by more than 8 times at pH 10. These results underscore the potential of the Fe3O4@LC-GLU-CALA system as a robust biocatalytic matrix with promising applications in biodiesel production, ester synthesis, and pharmaceutical manufacturing.

Multivariate mesoporous MOFs with regulatable hydrophilic/hydrophobic surfaces as a versatile platform for enzyme immobilization

Zeolitic imidazole frameworks materials-8 (ZIF-8), a member of the metal-organic framework (MOFs) series, have been extensively used as a host matrix for enzyme immobilization. However, the microporous structure and hydrophobicity, as well as the protonation of the precursor 2-methylimidazole (2-MeIm) of ZIF-8, remain considerable challenges in maintaining the activity of immobilized enzymes. Here, novel multivariate mesoporous MOFs (mMOFs) with regulatable hydrophilic/hydrophobic surfaces were designed by the multivariate competitive strategy and pore modification engineering. 3-Methyl-1H-1,2,4-triazole (3-MTZ) and 5-methyltetrazole (5-MTA) were employed to partially replace 2-MeIm. These were then combined with zinc sulfate heptahydrate (soft templates) to yield mMOFs in a methanol solution. As a proof-of-concept application, we used mMOFs as carriers for enzyme immobilization and investigated the properties of the immobilized enzymes. Benefiting from their mesoporous structure, hydrophilic surface, and improved microenvironment, multivariate mMOFs exhibit a strong ability to stabilize enzyme conformation and increase enzyme activity compared with traditional ZIF-8. Our study offers an avenue for the controllable preparation of well-designed MOF structures, which will further broaden the application opportunities of MOF materials for enzyme immobilization.

A molecular imprinting electrochemical sensor for detection of anticancer drug amsacrine

PurposeThe purpose of this study is to develop a molecular imprinting electrochemical sensor for the specific detection of the anticancer drug amsacrine. The sensor used a composite of bacterial cellulose (BC) and silver nanoparticles (AgNPs) as a platform for the immobilization of a molecularly imprinted polymer (MIP) film. The main objective was to enhance the electrochemical properties of the sensor and achieve a high level of selectivity and sensitivity toward amsacrine molecules in complex biological samples.Design/methodology/approachThe composite of BC-AgNPs was synthesized and characterized using FTIR, XRD and SEM techniques. The MIP film was molecularly imprinted to selectively bind amsacrine molecules. Electrochemical characterization, including cyclic voltammetry and electrochemical impedance spectroscopy, was performed to evaluate the modified electrode’s conductivity and electron transfer compared to the bare glassy carbon electrode (GCE). Differential pulse voltammetry was used for quantitative detection of amsacrine in the concentration range of 30–110 µM.FindingsThe developed molecular imprinting electrochemical sensor demonstrated significant improvements in conductivity and electron transfer compared to the bare GCE. The sensor exhibited a linear response to amsacrine concentrations between 30 and 110 µM, with a low limit of detection of 1.51 µM. The electrochemical response of the sensor showed remarkable changes before and after amsacrine binding, indicating the successful imprinting of amsacrine in the MIP film. The sensor displayed excellent selectivity for amsacrine in the presence of interfering substances, and it exhibited good stability and reproducibility.Originality/valueThis study presents a novel molecular imprinting electrochemical sensor design using a composite of BC and AgNPs as a platform for MIP film immobilization. The incorporation of BC-AgNPs improved the sensor’s electrochemical properties, leading to enhanced sensitivity and selectivity for amsacrine detection. The successful imprinting of amsacrine in the MIP film contributes to the sensor's specificity. The sensor's ability to detect amsacrine in a concentration range relevant to anticancer therapy and its excellent performance in complex sample matrices add significant value to the field of electrochemical sensing for pharmaceutical analysis.

The design of an RGD in situ sustained delivery system utilizing scallop byssal protein through genetic engineering

Although bioactive peptides enhancing bone healing have demonstrated effectiveness in treating bone defects, in vivo instability poses a challenge to their clinical application. Currently reported peptide delivery systems do not meet the demands of bone tissue repair regarding stability and peptide release efficacy. Herein, the self-assembling recombinant chimeric protein (Sbp5–2RGD) is developed by genetic engineering with cell adhesion peptide RGD as the targeted peptide and a newly discovered scallop byssal-derived protein Sbp5–2 that can assemble into wet stable films as the structural domain. In vitro studies show that the Sbp5–2RGD film exhibits excellent extensibility and biocompatibility. In vitro and in vivo degradation experiments demonstrate that the film remains stable due to the layer-by-layer degradation mode, resulting in sustained delivery of RGD in situ for up to 4 weeks. Consequently, the film can effectively promote osteogenesis, which accelerates bone defect healing and the implants osseointegration. Cell-level studies further show that the film up-regulates the expression of genes and proteins (ALP, OCN, OSX, OPN, RUNX2, VEGF) associated with osteogenesis and angiogenesis. Overall, this novel protein film represents an intelligent platform for peptide immobilization, protection, and release through its self-assembly, dense structure, and degradation mode, providing a therapeutic strategy for bone repair.

Alloyed Trimetallic Nanocomposite as an Efficient and Recyclable Solid Matrix for Ideonella sakaiensis Lipase Immobilization.

In this work, a trimetallic (Ni/Co/Zn) organic framework (tMOF), synthesized by a solvothermal method, was calcinated at 400 and 600 °C and the final products were used as a support for lipase immobilization. The material annealed at 400 °C (Ni-Co-Zn@400) had an improved surface area (66.01 m2/g) and pore volume (0.194 cm3/g), which showed the highest enzyme loading capacity (301 mg/g) with a specific activity of 0.196 U/mg, and could protect the enzyme against thermal denaturation at 65 °C. The optimal pH and temperature for the lipase were 8.0 and 45 °C but could tolerate pH levels 7.0-8.0 and temperatures 40-60 °C. Moreover, the immobilized enzyme (Ni-Co-Zn@Lipase, Ni-Co-Zn@400@Lipase, or Ni-Co-Zn@600@Lipase) could be recovered and reused for over seven cycles maintaining 80, 90, and 11% of its original activity and maintained a residual activity >90% after 40 storage days. The remarkable thermostability and storage stability of the immobilized lipase suggest that the rigid structure of the support acted as a protective shield against denaturation, while the improved pH tolerance toward the alkaline range indicates a shift in the ionization state attributed to unequal partitioning of hydroxyl and hydrogen ions within the microenvironment of the active site, suggesting that acidic residues may have been involved in forming an enzyme-support bond. The high enzyme loading capacity, specific activity, encouraging stability, and high recoverability of the tMOF@Lipase indicate that a multimetallic MOF could be a better platform for efficient enzyme immobilization.

Label-free electrochemical aptasensor for detection of Acinetobacter baumannii: Unveiling the kinetic behavior of reduced graphene oxide v/s graphene oxide

In this study, we introduce an electrochemical aptasensor for the sensitive detection of Acinetobacter baumannii. This method, based on impedance and current analysis, represents a first-of-its kind platform in the field of A. baumannii detection. We employed a screen-printed carbon electrode modified with both graphene oxide (GO) and reduced graphene oxide (rGO) as the immobilization platform, and the bacteria specific aptamer was employed as the biorecognition element. The aptamer selectively interacts with A. baumannii, causing a decrease in the peak current of the redox complex because the binding biomolecules on the electrode surface hinder the electron transfer of the redox probe. This initial point-of-care-compatible rGO electrochemical aptasensor demonstrated an exceptional sensing performance with an impressive limit of detection of 10 CFU/mL for detecting A. baumannii. This enhanced sensitivity can be attributed to several key factors including structural defects during reduction, increased surface area and improved electron transfer kinetics. The findings of this study highlight the potential of the developed aptasensor as a rapid and reliable detection platform for various pathogenic diseases. By further ensuring seamless integration into clinical practice, these label-free graphene-based aptasensors, with their π-π interactions and other favorable attributes, hold great promise for sensitive and specific pathogen detection, thereby contributing to advancements in healthcare diagnostics.

Carboxyethylsilanetriol-Coated Magnetic Nanoparticles as an Ultrasensitive Immunoplatform for Electrochemical Magnetosensing of Cotinine.

In the present study, an innovative and simple electrochemical magneto biosensor based on carboxyethylsilanetriol-modified iron oxide (Fe3O4) magnetic nanoparticles was designed for ultrasensitive and specific analysis of cotinine, an important marker of smoking. Anticotinine antibodies were covalently immobilized on carboxylic acid-modified magnetic nanoparticles, and the cotinine-specific magnetic nanoparticles created a specific surface on the working electrode surface. The use of magnetic nanoparticles as an immobilization platform for antibodies provided a large surface area for antibody attachment and increased sensitivity. In addition, the advantages of the new immobilization platform were reusing the working electrode numerous times, recording repeatable and reproducible signals, and reducing the necessary volume of biomolecules. The specific interaction between cotinine and cotinine-specific antibody-attached magnetic nanoparticles restricted the electron transfer of the redox probe and changed the impedimetric response of the electrode correlated to the concentration of cotinine. The magneto biosensor had a wide detection range (2-300 pg/mL), a low LOD (606 fg/mL), and an acceptable recovery (97.24-105.31%) in real samples. In addition, the current biosensor's measurement results were in good agreement with those found by the standard liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA) methods. These results showed that a simple impedimetric immunosensing platform was generated for the cotinine analysis.

Ultrasensitive Aptasensing Platform for the Detection of β-Amyloid-42 Peptide Based on MOF Containing Bimetallic Porphyrin Graphene Oxide and Gold Nanoparticles.

The prompt detection of diseases hinges on the accessibility and the capability to identify relevant biomarkers. The integration of aptamers and the incorporation of nanomaterials into signal transducers have not only expedited but also enhanced the development of nanoaptasensors, enabling heightened sensitivity and selectivity. Here, the bimetallic nickel-cobalt-porphyrin metal-organic framework ((Ni + Cu)TPyP MOF) is regarded as an electron mediator, immobilization platform for an Alzheimer aptamer and to increase the electrochemical signal for the detection of the main biomarker of Alzheimer's disease (AD), amyloid β (Aβ-42). Furthermore, the ((Ni + Cu)TPyP MOF) was combined with reduced graphene oxide (rGO) and gold nanoparticles (AuNPs), on a gold electrode (GE) to provide an efficient interface for immobilizing aptamer strands. Concurrently, the incorporation of rGO and AuNPs imparts enhanced electrical conductivity and efficacious catalytic activity, establishing them as adept electrochemical indicators. Owing to the superior excellent electrical conductivity of rGO and AuNPs, coupled with the presence of ample mesoporous channels and numerous Ni and Cu metal sites within (Ni + Cu)TPyP MOF, this nanostructure with abundant functional groups is proficient in immobilizing a substantial quantity of aptamer. These interactions are achieved through robust π-π stacking and electrostatic interactions, alongside the high affinity between the thiol group of the aptamer and AuNPs concurrently. The as-prepared ternary (Au@(Ni + Cu)TPyP MOF/rGO) nanostructure electrode exhibited an enhancement in its electrochemically active surface area of about 7 times, compared with the bare electrode and the Aβ-42 redox process is highly accelerated, so the peak currents are significantly higher than those obtained with bare GE substrate. Under the optimized conditions, the designed aptasensor had the quantitative detection of Aβ-42 with a low detection limit of 48.6 fg mL-1 within the linear range of 0.05 pg mL-1 to 5 ng mL-1 by differential pulse voltammetry (DPV), accompanied by precise reproducibility, satisfactory stability (95.6% of the initial activity after 10 days), and minimal impact of interfering agents. Recorded results in human blood plasma demonstrated the high efficacy of porphyrin MOF system sensing even in the clinical matrix. The great performance of this aptasensor indicates that our new design of Au@(Ni + Cu)TPyP MOF/rGO nanostructure provides more opportunities for the detection of chemical signals in early diagnosis of Alzheimer's disease.

Double-Network structure chemically programmed DNA Cryogel: Enzyme immobilization platform with enhanced catalytic activity for portable biosensing

Enzyme-based hydrogels have conferred multiple benefits, particularly for disease diagnosis and environmental monitoring due to their pronounced biocompatibility, ability to load cargo, and optical properties. However, traditional hydrogels suffer from some significant limitations, such as enzyme leakage and catalytic activity decreased, restricting the extent to which hydrogels can be beneficially used. Herein, chemically programmed DNA-DNA interactions were designed as a promising solution for enzyme immobilization that substantially maintains biological functionality with remarkable leakage reduction. Additionally, their responsiveness and mechanical properties are dramatically enhanced after adjusting the porous structure of hydrogels via freezing. As a proof-of-concept, a DNA cryogel with a hierarchical structure was developed with adjustable distance among cascade enzymes, increasing the local concentration of intermediates and reducing their mass transfer energy barriers for direct glucose monitoring and glutathione (GSH) sensing. Moreover, after being integrated with a smartphone system, a universal sensing platform (immunosensor) was fabricated for rapid detection of environmental chemicals (i.e., bisphenol A) without expensive instruments and sophisticated procedures. This strategy provided some novel insights on hydrogel- based bioassays, implying their great potentials for conducting onsite analysis against various targets.

Improved L-Asparaginase Properties and Reusability by Immobilization onto Functionalized Carbon Xerogels.

Enzyme immobilization can offer a range of significant advantages, including reusability, and increased selectivity, stability, and activity. In this work, a central composite design (CCD) of experiments and response surface methodology (RSM) were used to study, for the first time, the L-asparaginase (ASNase) immobilization onto functionalized carbon xerogels (CXs). The best results were achieved using CXs obtained by hydrothermal oxidation with nitric acid and subsequent heat treatment in a nitrogen flow at 600 °C (CX-OX-600). Under the optimal conditions (81 min of contact time, pH 6.2 and 0.36 g/L of ASNase), an immobilization yield (IY) of 100 % and relative recovered activity (RRA) of 103 % were achieved. The kinetic parameters obtained also indicate a 1.25-fold increase in the affinity of ASNase towards the substrate after immobilization. Moreover, the immobilized enzyme retained 97 % of its initial activity after 6 consecutive reaction cycles. All these outcomes confirm the promising properties of functionalized CXs as support for ASNase, bringing new insights into the development of an efficient and stable immobilization platform for use in the pharmaceutical industry, food industry, and biosensors.