Construction of Covalent Organic Framework Capsule-Based Nanoreactor for Sensitive Glucose Detection.

Enzyme immobilization is critical to boosting its application in various areas. Covalent organic frameworks (COFs) are ideal hosts for enzyme immobilization due to their porous and predesignable structures. Nevertheless, the construction of COFs-based enzyme immobilization systems with high activity via existing immobilization methods (including covalent linkages and channel entrapment) remains a considerable challenge. Herein, a versatile approach was introduced to encapsulate enzymes within hollow COF capsule (named enzyme@COF) using metal-organic frameworks (including ZPF-1(C8H11N4O4.5Zn), ZIF-8(C8H10N4Zn), and ZIF-90(C8H6N4O2Zn)) as sacrificial templates. The obtained porous COF capsule could not only facilitate the efficient mass transfer of enzymatic reactions but also protect enzymes against the incompatible conditions, resulting in enhanced activity and stability of the encapsulated enzymes. Moreover, this approach offered an opportunity to spatially organize multienzymes in COF capsule to construct enzyme cascade system. For instance, glucose oxidase (GOx) and cytochrome c (Cyt c) were coencapsulated within COF capsule to construct GOx-Cyt c cascade. The integration of GOx and Cyt c within COF capsule achieved ∼1.6-fold improvement in catalytic activity than that of free enzymes and the resultant GOx-Cyt c@COF was successfully adopted as a nanoreactor for the sensitive determination of glucose in serum. This work provided a new insight into the design of COFs-based enzyme immobilization systems.

Construction of Donor-Acceptor Heteroporous Covalent Organic Frameworks as Photoregulated Oxidase-like Nanozymes for Sensing Signal Amplification.

Nanomaterials with enzyme-like characteristics (called nanozymes) show their extreme potentials as alternatives to natural enzymes. Covalent organic frameworks (COFs) as metal-free nanozymes have attracted huge attention for catalytic applications due to their flexible molecular design and synthetic strategies and conjugated, porous, and chemically stable architectures. Designing high-performance two-dimensional (2D) porous COF materials embedded with functional building units for modulating nanozymes' catalytic activity is of immense importance in contemporary research. The proper combination of donor-acceptor (D-A) fragments within a porous COF skeleton is an effective strategy to decrease the band gap and provide a strong charge-transfer pathway for highly effective charge separation. Herein, two donor-acceptor heteroporous COFs using an electron-deficient 4,4'-(thiazolo[5,4-d]thiazole-2,5-diyl)dibenzaldehyde (Tz) unit or 4,4'-(benzo[c][1,2,5]thiadiazole-4,7-diyl)dibenzaldehyde (Td) unit and electron-rich tetrakis(4-aminophenyl)ethane (ETTA) linkers were presented. The resulting crystalline and heteroporous COFs showed outstanding oxidase-like activity under light irradiation, which can catalyze the oxidation of typical substrates and corresponding evolution in color and absorption. The light-activatable ETTA-Tz COF with prominent oxidase-like activity can serve as a colorimetric probe for quantitative detection of sulfide ions with a linear range of 1-50 µM and a detection limit of 0.27 µM within 3 min. The colorimetric approach could also be used for sulfide ion detection in human serum samples. The research demonstrated the future potential of D-A motifs within fully conjugated COFs to obtain excellent mimic enzyme activity.

Nanoscale Covalent Organic Frameworks with Donor-Acceptor Structures as Highly Efficient Light-Responsive Oxidase-like Mimics for Colorimetric Detection of Glutathione.

Although organic artificial enzymes have been reported as biomimetic oxidation catalysts and are widely used for colorimetric biosensors, developing organic artificial enzymes with high enzymatic activity is still a challenge. Two-dimensional (2D) covalent organic frameworks (COFs) have shown superior potential in biocatalysts because of their periodic π-π arrays, tunable pore size and structure, large surface area, and thermal stability. The interconnection of electron acceptor and donor building blocks in the 2D conjugated COF skeleton can lead to narrower band gaps and efficient charge separation and transportation and thus is helpful to improve catalytic activity. Herein, a donor-acceptor 2D COF was synthesized using tetrakis(4-aminophenyl)pyrene (Py) as an electron donor and thieno[3,2-b]thiophene-2,5-dicarbaldehyde (TT) as an electron acceptor. Under visible light irradiation, the donor-acceptor 2D COF exhibited superior enzymatic catalytic activity, which could catalyze the oxidation of chromogenic substrates such as 3,3',5,5'-tetramethylbenzidine (TMB) by the formation of superoxide radicals and holes. Based on the above property, the photoactivated donor-acceptor 2D COF with enzyme-like catalytic properties was designed as a robust colorimetric probe for cheap, highly sensitive, and rapid colorimetric detection of glutathione (GSH); the corresponding linear range of GSH was 0.4-60 µM, and the limit of detection was 0.225 µM. This study not only presents the construction of COF-based light-activated nanozymes for environmentally friendly colorimetric detection of GSH but also provides a smart strategy for improving nanozyme activity.

Room-temperature controllable synthesis of hierarchically flower-like hollow covalent organic frameworks for brain natriuretic peptide enrichment.

A facile strategy was introduced for room-temperature controllable synthesis of hierarchically flower-like hollow COFs (FHF-COFs). Furthermore, the universality for synthesis of the HFH-COFs was validated by altering the building units. Inspired by the unique morphology, extremely large surface area and good chemical stability, HFH-COFs could serve as an attractive adsorption probe by loading with gold nanoparticles and be applied to enrichment of brain natriuretic peptide from human serum. This work opens up a whole new approach for controllable synthesis of the HFH-COFs at room temperature and expands the application of COFs as a promising enrichment probe for complex biological samples.

Facile synthesis of bifunctional polymer monolithic column for tunable and specific capture of glycoproteins and phosphoproteins.

Efficiently tunable capture of the glycosylated/phosphorylated proteins is critical to meet the need of in-depth glycoproteome and phosphoproteome studies. Reported here is a new bifunctional polymer monolithic column by introducing benzeneboronic acid and phosphonic acid onto monolithic column (denoted as poly (EDMA-co-VPBA-co-VPA) monolith) for tunable and specific enrichment of glycoproteins and phosphoproteins via switching different mobile phases. Based on boronate affinity and immobilized metal affinity, the as-prepared poly (EDMA-co-VPBA-co-VPA) monolith exhibited superior performance in selective separation of small molecules and biomacromolecules containing cis-diol/phosphate groups or not. And the frontal chromatography analysis showed that the binding capacity of the poly (EDMA-co-VPBA-co-VPA) monolith towards horseradish peroxidase (HRP, glycoprotein) or ß-casein (phosphoprotein) is four-fold higher than that of bovine serum albumin (BSA, non-glycosylated/phosphorylated protein). Furthermore, combined with mass spectrometry identification, the successful application in specific enrichment of glycopeptides/phosphopeptides from tryptic digests of HRP/ß-casein and direct capture of low abundant endogenous phosphopeptides from human serum proved great practicability in complex samples. This study provides a novel insight for fabricating the monolithic columns with multifunctionalization to facilitate further post-translational modification (PTM)-proteomics development.