Enzymatic Nanoreactors Research Articles

Enhanced Catalytic Stability of Laccase Immobilized on Copper Oxide Nanoparticles

We describe the design and catalytic activity of enzymatic nanoreactors based on copper oxide nanoparticles surface‐functionalized with laccase (CuO‐Lac) from Coriolopsis gallica. Transmission electron micrographs show complete laccase coverage on the nanoparticle surfaces. In addition, the catalytic rate (kcat) of the immobilized laccase is two‐fold higher at an optimized pH, when compared with free laccase, while the affinity constant (KM) is not significantly affected. Importantly, the total turnover number (TNN) of the CuO‐Lac nanoreactors reaches 358 ± 9 × 106 mol ABTS/mol laccase, which is four times higher than the TTN obtained for the free enzyme (88 ± 3 × 106). Thus, our nanoreactors exhibit a significant improvement in laccase catalytic performance. Inductively coupled plasma mass spectrometry was used to confirm the loss of copper ions from free laccase during the catalytic cycle, suggesting that the CuO nanoparticles act as a copper ion source that prevents enzyme inactivation. This work contributes to an improved understanding of the CuO‐laccase interface and confirms that CuO nanoparticles serve as copper sources for laccase during catalysis.

Magnetic biocatalytic nanoreactors based on graphene oxide with graded reduction degrees for the enzymatic synthesis of phytosterol esters

As we know, the carriers of enzymatic nanoreactors with different amphipathy have significant influence on the interfacial enzyme activities and its performance. In this study, the magnetic graphene oxide (GO) enzyme carriers were constructed by assembling graphene oxide with different reduction degrees (rGO(x)) and polyethyleneimine (PEI) modified nano-flowery Fe3O4 into composite materials through electrostatic interactions, and x here represents the reduction reaction hours of GO. The SEM, TEM, contact angles and BET tests qualified the ideal enzyme carrier, PEI-Fe3O4-rGO(12), by adsorption-crosslinking method, the Candida rugosa lipase (CRL) was immobilized to obtain the lipase nanoreactor, PEI-Fe3O4-rGO(12)@CRL. The nano-sheet and nano-flower structures provided a larger specific surface area and suitable pore size, and the CRL had higher protein load on the carrier material (163.11 mg/g). In addition, using hydrolyzed p-nitrophenol palmitate as the model, the PEI-Fe3O4-rGO(12)@CRL can still show 76.5 % residual activity after 8 repeated uses. At the same time, the immobilized lipase showed good catalytic performance in the biosynthesis of phytosterol esters. The optimal esterification conditions were determined by single factor experiment (isooctane, sterol concentration 150 μmol/mL, alkyd molar ratio 1:2, enzyme dosage 120 mg, 26 h, 55 °C, the esterification rate was 76.24 %). Therefore, the optimal biocatalyst PEI-Fe3O4-rGO(12)@CRL may has good performance in the biosynthesis of other novel foods.

Stable and permeable polyion complex vesicles designed as enzymatic nanoreactors.

Polymeric vesicles are perspective vehicles for fabricating enzymatic nanoreactors towards diverse biomedical and catalytic applications, yet the design of stable and permeable vesicles remains challenging. Herein, we developed polyion complex (PIC) vesicles featuring high stability and a permeable membrane for adequate enzyme loading and activation. Our design relies on co-assembly of an anionic diblock copolymer (PSS96-b-PEO113) with cationic branched poly(ethylenimine) (PEI). The polymer combination endows strong electrostatic interaction between the PSS and PEI building blocks, so their assembly can be implemented at a high salt concentration (500 mM NaCl), under which the charge interaction of the enzyme-polymer is inhibited. This control realizes the successful and safe loading of enzymes associated with the formation of stable PIC vesicles with an intrinsic permeable membrane that is favourable for enhancing enzymatic activity. The control factors for vesicle formation and enzyme loading were investigated, and the general application of loading different enzymes for cascade reaction was validated as well. Our study reveals that proper design and combination of polyelectrolytes is a facile strategy for fabricating stable and permeable polymeric PIC vesicles, which exhibit clear advantages for loading and activating enzymes, consequently boosting their diverse applications as enzymatic nanoreactors.

Geometric and defects engineering collaboration for enhanced cascade enzymatic nanoreactors

Geometric and defects engineering collaboration for enhanced cascade enzymatic nanoreactors

Nanoparticle Concentration as an Important Parameter for Characterization of Dispersion and Its Applications in Biomedicine

Currently, there are problems to standardize methods for determining the concentration of nanoparticles and creation of etalon materials for calibrating measured concentrations. Accurate determination of nanoparticle concentration is necessary to assess the maximum dose of administered nanotherapeutics for diagnostics and therapy in vivo, to determine the order of reaction in enzymatic nanoreactors. In addition, this parameter determines biological effects, such as the formation of a protein corona on the outer surface of nanoparticles that precedes nanoparticles’ absorption and internalization in cells. This review discusses the most common methods for determining the concentration of nanoparticles based on direct visualization, using microscopy, light absorption or light scattering, direct counting of nanoparticles, and gravimetry. Results may differ from one method to the other. Thus, the use of a combination of several methods provides more reliable results. The advantages, disadvantages and ways to improve accuracy of results are also presented.

Glucose oxidase virus-based nanoreactors for smart breast cancer therapy.

Breast cancer is the most common malignant tumor disease and the leading cause of female mortality. The evolution of nanomaterials science opens the opportunity to improve traditional cancer therapies, enhancing therapy efficiency and reducing side effects. Herein, protein cages conceived as enzymatic nanoreactors were designed and produced by using virus-like nanoparticles (VLPs) from Brome mosaic virus (BMV) and containing the catalytic activity of glucose oxidase (GOx) enzyme. The GOx enzyme was encapsulated into the BMV capsid (VLP-GOx), and the resulting enzymatic nanoreactors were coated with human serum albumin (VLP-GOx@HSA) for breast tumor cell targeting. The effect of the synthesized GOx nanoreactors on breast tumor cell lines was studied in vitro. Both nanoreactor preparations VLP-GOx and VLP-GOx@HSA showed to be highly cytotoxic for breast tumor cell cultures. Cytotoxicity for human embryonic kidney cells was also found. The monitoring of nanoreactor treatment on triple-negative breast cancer cells showed an evident production of oxygen by the catalase antioxidant enzyme induced by the high production of hydrogen peroxide from GOx activity. The nanoreactors containing GOx activity are entirely suitable for cytotoxicity generation in tumor cells. The HSA functionalization of the VLP-GOx nanoreactors, a strategy designed for selective cancer targeting, showed no improvement in the cytotoxic effect. The GOx containing enzymatic nanoreactors seems to be an interesting alternative to improve the current cancer therapy. In vivo studies are ongoing to reinforce the effectiveness of this treatment strategy.

Targeted Enzymatic VLP-Nanoreactors with β-Glucocerebrosidase Activity as Potential Enzyme Replacement Therapy for Gaucher's Disease.

Gaucher disease is a genetic disorder and the most common lysosomal disease caused by the deficiency of enzyme β-glucocerebrosidase (GCase). Although enzyme replacement therapy (ERT) is successfully applied using mannose-exposed conjugated glucocerebrosidase, the lower stability of the enzyme in blood demands periodic intravenous administration that adds to the high cost of treatment. In this work, the enzyme β-glucocerebrosidase was encapsulated inside virus-like nanoparticles (VLPs) from brome mosaic virus (BMV), and their surface was functionalized with mannose groups for targeting to macrophages. The VLP nanoreactors showed significant GCase catalytic activity. Moreover, the Michaelis-Menten constants for the free GCase enzyme (KM =0.29 mM) and the functionalized nanoreactors (KM =0.32 mM) were similar even after chemical modification. Importantly, the stability of enzymes under physiological conditions (pH 7.4, 37 °C) was enhanced by ≈11-fold after encapsulation; this is beneficial for obtaining a higher blood circulation half-life, which may decrease the cost of therapy by reducing the requirement of multiple intravenous injections. Finally, the mannose receptor targeted enzymatic nanoreactors showed enhanced internalization into macrophage cells. Thus, the catalytic activity and cell targeting suggest the potential of these nanoreactors in ERT of Gaucher's disease.

Kinetic Processes in Enzymatic Nanoreactors for In Vivo Detoxification.

Enzymatic nanoreactors are enzyme-encapsulated nanobodies that are capable of performing biosynthetic or catabolic reactions. For this paper, we focused on therapeutic enzyme nanoreactors for the neutralization of toxicants, paying special attention to the inactivation of organophosphorus compounds (OP). Therapeutic enzymes that are capable of detoxifying OPs are known as bioscavengers. The encapsulation of injectable bioscavengers by nanoparticles was first used to prevent fast clearance and the immune response to heterologous enzymes. The aim of enzyme nanoreactors is also to provide a high concentration of the reactive enzyme in stable nanocontainers. Under these conditions, the detoxification reaction takes place inside the compartment, where the enzyme concentration is much higher than in the toxicant diffusing across the nanoreactor membrane. Thus, the determination of the concentration of the encapsulated enzyme is an important issue in nanoreactor biotechnology. The implications of second-order reaction conditions, the nanoreactor’s permeability in terms of substrates, and the reaction products and their possible osmotic, viscosity, and crowding effects are also examined.

Block Copolymer Vesicles with Tunable Membrane Thicknesses and Compositions Prepared by Aqueous Seeded Photoinitiated Polymerization-Induced Self-Assembly at Room Temperature.

Block copolymer vesicles with diverse functionalities and intrinsic hollow structures have received considerable attention due to their broad applications in biomedical fields, including drug delivery, bioimaging, theranostics, gene therapy, etc. However, efficient preparation of block copolymer vesicles with tunable membrane thicknesses and compositions under mild conditions is still a challenge. Herein, we report an aqueous seeded photoinitiated polymerization-induced self-assembly (photo-PISA) for the precise preparation of block copolymer vesicles at room temperature. By changing the total degree of polymerization (DP) of the hydrophobic block in seeded photo-PISA, one can easily tune the membrane thickness without compromising the morphology of vesicles. Moreover, by adding different comonomers such as hydrophobic monomers, hydrophilic monomers, and cross-linkers into seeded photo-PISA, vesicles with different compositions could be prepared without compromising the morphology and colloidal stability. Polymerization kinetics show that seeded photo-PISA can skip the step of in situ self-assembly with a short homogeneous polymerization stage being observed. To demonstrate potential biological applications, enzymatic nanoreactors were constructed by loading horseradish peroxidase (HRP) inside vesicles via seeded photo-PISA. The enzymatic properties of these nanoreactors could be easily regulated by changing the membrane thickness and hydrophobicity. It is expected that this method can provide a facile platform for the precise preparation of block copolymer vesicles that may find applications in different fields.

Concentrically Encapsulated Dual-Enzyme Capsules for Synergistic Metabolic Disorder Redressing and Cytotoxic Intermediates Scavenging.

Enzyme therapy has important implications for the treatment of metabolic disorders and biological detoxification. It remains challenging to prepare enzymatic nanoreactors with high therapeutic efficiency and low emission of cytotoxic reaction intermediates. Here, we propose a novel strategy for the preparation of enzymes-loaded polypeptide microcapsules (EPM) with concentrically encapsulated enzymes to achieve higher cascade reaction rates and minimal emission of cytotoxic intermediates. Mesoporous silica spheres (MSS) are used as a highly porous matrix to efficiently load a therapeutic enzyme (glucose oxidase, GOx), and a layer-by-layer (LbL) assembly strategy is employed to assemble the scavenging enzyme (catalase) and polyelectrolyte multilayers on the MSS surface. After removal of the MSS, a concentrically encapsulated EPM is obtained with the therapeutic enzyme encapsulated inside the capsule, and the scavenging enzyme immobilized in the polypeptide multilayer shell. Performance of the concentrically encapsulated GOx-catalase capsules is investigated for synergistic glucose metabolism disturbance correction and cytotoxic intermediate H2O2 clearance. The results show that the EPM can simultaneously achieve 99% H2O2 clearance and doubled glucose consumption rate. This strategy can be extended to the preparation of other dual- or multi-enzyme therapeutic nanoreactors, showing great promise in the treatment of metabolic disorders.

Effect of Functional Group on the Catalytic Activity of Lipase B from Candida antarctica Immobilized in a Silica-Reinforced Pluronic F127/α-Cyclodextrin Hydrogel

Surface modification plays a key role in the fabrication of highly active and stable enzymatic nanoreactors. In this study, we report for the first time the effect of various functional groups (epoxy, amine, trimethyl, and hexadecyl) on the catalytic performance of lipase B from Candida antarctica (CALB) incorporated within a monolithic supramolecular hydrogel with multiscale pore architecture. The supramolecular hydrogel formed by host-guest interactions between α-cyclodextrin (α-CD) and Pluronic F127 was first silicified to provide a hierarchically porous material whose surface was further modified with different organosilanes permitting both covalent anchoring and interfacial activation of CALB. The catalytic activity of nanoreactors was evaluated in the liquid phase cascade oxidation of 2,5-diformylfuran (DFF) to 2,5-furandicarboxylic acid (FDCA) under mild conditions. Results showed that high FDCA yields and high efficiency conversion of DFF could be correlated with the ability of epoxy and amine moieties to keep CALB attached to the carrier, while the trimethyl and hexadecyl groups could provide a suitable hydrophobic-hydrophilic interface for the interfacial activation of lipase. Cationic cross-linked β-CD was also evaluated as an enzyme-stabilizing agent and was found to provide beneficial effects in the operational stability of the biocatalyst. These supramolecular silicified hydrogel monoliths with hierarchical porosity may be used as promising nanoreactors to provide easier enzyme recovery in other biocatalytic continuous flow processes.

Rapid sample preparation workflow based on enzymatic nanoreactors for potential serum biomarker discovery in pancreatic cancer

Mass spectrometry (MS)-based proteomics have been extensively applied in clinical practice to discover potential protein and peptide biomarkers. However, the traditional sample pretreatment workflow remains labor-intensive and time-consuming, which limits the application of MS-based proteomic biomarker discovery studies in a high throughput manner. In the current work, we improved the previously reported procedure of the simple and rapid sample preparation methods (RSP) by introducing macroporous ordered siliceous foams (MOSF), namely RSP-MOSF. With the aid of MOSF, we further reduced the digestion time to 10 min, facilitating the whole sample handling process within 30 min. Combining with 30 min direct data independent acquisition (DIA) of LC-MS/MS, we accomplished a serum sample analysis in 1 h. Comparing with the RSP method, the performance of protein and peptide identification, quantitation, as well as the reproducibility of RSP-MOSF is comparable or even outperformed the RSP method. We further applied this workflow to analyze serum samples for potential candidate biomarker discovery of pancreatic cancer. Overall, 576 serum proteins were detected with 41 proteins significantly changed, which could serve as potential biomarkers for pancreatic cancer. Additionally, we evaluated the performance of RSP-MOSF method in a 96-well plate format which demonstrated an excellent reproducibility of the analysis. These results indicated that RSP-MOSF method had the potential to be applied on an automatic platform for further scaled analysis.

UV-light-actuated in-situ preparation of paper@ZnCd quantum dots for paper-based enzymatic nanoreactors

Quantum dots (QDs) have been widely applied in the analytical field including sensitive fluorescent assays on paper-based devices. A facile in-situ synthesis of QDs after a simple reagent deposition on paper could enable a rapid low-cost fabrication of QD–modified paper with a variety of properties and purposes such as enzymatic nanoreactors. Herein, for the first time, fluorescent ZnCd QDs were prepared by in-situ synthesis in the paper matrix using thiol-containing precursors and irradiation by UV light. A successful creation of the immobilized ZnCd QDs on paper (paper@ZnCd QDs) was monitored by their intrinsic fluorescence while their peroxidase mimetic activity was evaluated by a catalytic reaction between H2O2 and a substrate (3,3′,5,5′-tetramethylbenzidine, TMB) producing a blue coloured product of charged transfer complex of TMB (oxTMB). From three thiol precursors investigated, the insulin precursor provided a greater activity than glutathione (GSH) and both considerably larger than BSA. Finally, a QDs precursor mixture deposition onto the paper matrix was evaluated by the peroxidase-mimetic activity, which was comparable with a reference method. The in-situ preparation of paper@ZnCd QDs is simple, rapid and ‘green’, with potential in biomedical applications primarily as fluorescence imaging agent and enzyme mimetic paper-based nanoreactors.

Construction of enzymatic nanoreactors with high catalytic activity in millifluidic systems for cancer therapy

Enzymatic nanoreactors have shown great potential in cancer therapy due to their delighted anticancer results. Synthesizing biocompatible and highly active nanoreactors on a large scale is demanded in clinic translation. Herein, the therapeutic enzymatic nanoreactors with the structure of biocompatible micromaterials covered on enzymes were constructed in the millifluidic device. The results indicated that the nanoreactors were successfully synthesized in the droplet-flow millifluidic device for the first time and had high bio-safety even without post-processing. The nanoreactors could be internalized by cancer cells and possessed long blood circulation time in vivo (t1/2 = 25 h). Moreover, mild synthetic conditions and minimized synthesis pathway empowered nanoreactors with favorable catalytic activity about 96.5% ± 3.9% of native enzymes. Furthermore, enzymatic nanoreactors loaded with glucose oxidase presented a strong anticancer effect and low systemic toxicity. In summary, the assay provides a facile and promising strategy for synthesizing highly active nanoreactors on a large scale.

Enzymatically Active DNA-Protein Nanogels with Tunable Cross-Linking Density.

Hybrid DNA-protein nanogels represent potential protein vectors and enzymatic nanoreactors for modern biotechnology. Here, we describe a new, easy, and robust method for preparation of tunable DNA-protein nanogels with controllable size and density. For this purpose, polymerase chain reaction is used to prepare highly biotinylated DNA as a soft biopolymeric backbone, which can be efficiently cross-linked via streptavidin-biotin binding. This approach enables us to control both the density and size of the resulting nanogels not only by adjusting the amount of the cross-linking streptavidin but also by using different rates of DNA biotinylation. This gives DNA-streptavidin nanogels with the size ranging from 80 nm, for the most compact state, to up to 200 nm. Furthermore, using streptavidin-enzyme conjugates allows the straightforward one-pot incorporation of enzymes during the preparation of the nanogels. Monoenzymatic and multienzymatic nanogels have been obtained in this manner, and their catalytic activities have been characterized. All tested enzymes (alkaline phosphatase (AP), horseradish peroxidase (HRP), and β-galactosidase (βGal)), incorporated individually or in a coupled manner (glucose oxidase (GOx)-HRP cascade), were shown to remain functional. The activities of AP and βGal were unchanged while that of HRP was slightly improved inside the nanogels. We demonstrate that, for HRP, it is not the DNA-to-enzyme ratio but the physical density of the functionalized DNA nanogels that is responsible for the improvement of its enzymatic activity.

Highly sensitive and portable aptasensor by using enzymatic nanoreactors as labels

Due to the extremely low concentration of many small molecular targets, improving the sensitivity of aptasensors is urgently needed. Herein, a highly sensitive and portable biosensing platform of Fe3O4@Au-Apt/DMSNs-I-cDNA is designed on magnetic Fe3O4@Au by using enzymatic nanoreactors as signal labels. This signal label is prepared by encapsulating invertase into dendritic mesoporous silica nanoparticles (DMSNs-I), resulting in large enzyme load amount (407.3 ± 28 mg g−1) and high bioactivity recovery (78.56%). Therefore, DMSNs-I can efficiently convert sucrose into glucose with personal glucose meter (PGM) as readout. Under PGM assay mode, this sensing platform achieves accurate recognition and sensitive detection of aflatoxin B1 (AFB1) and bisphenol A (BPA) with low detection limits of 0.74 pg mL−1 and 0.094 pM (S/N = 3), respectively. Compared with the single invertase labelled aptasensor, the DMSNs-I labelled AFB1 aptasensor achieves ~3.19 times enhancement of sensitivity and 2 orders of magnitude lower in the detection limit. In addition, the proposed aptasensors are applied to detect AFB1 and BPA in complex samples with the acceptable recovery values. They also exhibited good selectivity and high stability, indicating the potential of the Fe3O4@Au-Apt/DMSNs-I-cDNA biosensing platform in constructing highly sensitive aptasensors for detecting small molecules.

Colorimetric biosensing of organophosphate pesticides using enzymatic nanoreactor built on zeolitic imdiazolate-8

A nanoreactor (NR) was synthesized by encapsulating acetylcholinesterase (AChE) in zeolitic imidazolate framework (ZIF)-8 (AChE@ZIF-8 NR). The catalytic activity of the NR was tested for the hydrolysis of acetylthiocholine chloride (ATCh) based on the Ellman’s assay to yield a characteristic yellow color at a maximum wavelength (λmax) of 412 nm. The catalytic activity of AChE@ZIF-8 NR showcased noticeable enhancements in maximum reaction rate (νmax: 1.25 × 10−12 kat) and low Michaelis-Menton constant (Km: 0.05 mM (10 ppm)). The NR-based hydrolysis of ATCh was inhibited in the presence of each of all seven types of organophosphate pesticides (malathion (MAL), methyl parathion (MP), glyphosate (GLY), glufosinate (GLU), chlorpyrifos (CPF), dichlorvos (DCV), and paraoxon (PAR)) selected in this study. The limit of detection (LOD) values for GLY, GLU, and MAL were estimated almost identically as 10−5 ppb. However, their limit of quantification (LOQ) was slightly distinguished as 0.05, 0.01, and 0.05 ppb, respectively. The NR retained 46% of its activity over the course of 5 cycles of sequential activation and MAL-induced inhibition. In comparison to the other reported studies, our approach provides a rapid means for sensing of OPs with appreciable stability, enhanced recyclability, and facile observation.

Mesoporous Silica as Sorbents and Enzymatic Nanoreactors for Microbial Membrane Proteomics.

The membrane proteins of microbes are at the forefront of host and parasite interactions. Having a general view of the functions of microbial membrane proteins is vital for many biomedical studies on microbiota. Nevertheless, due to the strong hydrophobicity and low concentration of membrane proteins, it is hard to efficiently enrich and digest the proteins for mass spectrometry analysis. Herein, we design an enzymatic nanoreactor for the digestion of membrane proteins using methylated well-ordered hexagonal mesoporous silica (Met-SBA-15). The material can efficiently extract hydrophobic membrane proteins and host the proteolysis in nanopores. The performance of the enzymatic nanoreactor is first demonstrated using standard hydrophobic proteins and then validated using membrane proteins extracted from Escherichia coli (E. coli) or a mixed bacterial sample of eight strains. Using the nanoreactor, 431 membrane proteins are identified from E. coli, accounting for 38.5% of all membrane proteins of the species, which is much more than that by the widely used in-solution digestion protocol. From the mixed bacterial sample of eight strains, 1395 membrane proteins are identified using the nanoreactor. On the contrary, the traditional in-solution proteolysis workflow only leads to the identification of 477 membrane proteins, demonstrating that the Met-SBA-15 can be offered as an excellent tool for microbial membrane proteome research and is expected to be used in human microbiota studies, e.g. host-microbe interactions.

Virus-Based Nanoreactors with GALT Activity for Classic Galactosemia Therapy.

Enzymatic nanoreactors were obtained by galactose-1-phosphate uridylyl-transferase (GALT) encapsulation into plant virus capsids by a molecular self-assembly strategy. The aim of this work was to produce virus-like nanoparticles containing GALT for an enzyme-replacement therapy for classic galactosemia. The encapsulation efficiency and the catalytic constants of bio-nanoreactors were determined by using different GALT and virus coat protein ratios. The substrate affinity of nanoreactors was slightly lower than that of the free enzyme; the activity rate was 16 % of the GALT free enzyme. The enzymatic nanoreactors without functionalization were internalized into different cell lines including fibroblast and kidney cells, but especially into hepatocytes. The enzymatic nanoreactors are an innovative enzyme preparation with potential use for the treatment of classic galactosemia.

Metal-organic frameworks (MOFs) for biopreservation: From biomacromolecules, living organisms to biological devices

Preservation of biological entities (e.g. DNA, enzymes, viruses, microorganisms, biochips, etc.) has presented great potential in industrial catalysis and biomedical engineering due to their enhanced structural stability and biological functionality. However, technology gaps still exist for achieving high encapsulation efficiency with low performance deterioration to provide a universal preservation method for biological entities. Metal-organic frameworks (MOFs) have been demonstrated as promising platforms for encapsulation of different bioentities for their structural diversity, versatile tailorability as well as ultrahigh loading capacity. This review comprehensively summarizes the recent progress for encapsulation of bioentity within MOF ([email protected]) via post-synthetic infiltration or in situ encapsulation, and classifies into three sections: biomacromolecules, viruses and living organisms, and biological devices. MOF coating can not only behavior as protective porous exoskeletons to develop enzymatic nanoreactors that allow the access of small substrates through the narrow channel. The emergence of bio-decomposable MOFs can be also employed for preserving functional bioentities (e.g. bio-based therapeutics, clinical biomarkers, cells, viruses, diagnostic biochips, etc.) that can retain their original activities after MOF being eluted. Predictably, the MOF preservation technique might bring in the “technical innovation” in industrial and/or biomedical applications.