We report a novel turn-on sensing probe for the detection of butyrylcholinesterase activity in human serum using a fluorophore with high binding affinity for HSA.

Enzyme immobilization is very important for diverse enzyme applications. Particularly, there is a growing need for coimmobilization of multiple enzymes for biosensing and synthetic applications. However, it is still challenging to coimmobilize two enzymes with desirable features, including high immobilization yield, retention of enzymatic activity, and low leaching. In this study, we demonstrated that a pluronic-based nanocarrier (PNC) can be an encapsulation platform for immobilization of various single enzymes. Since the PNC is temperature-sensitive, a simple temperature change from 4 to 37 °C led to a substantial size reduction and enzyme encapsulation. All six enzymes tested were encapsulated by the PNC in high yield (∼90%) with the retained enzymatic activity (>95%). The leaching of encapsulated enzymes was very minimal (<0.13% for 2 weeks). Then, we demonstrated that the PNC can efficiently coencapsulate two enzymes, formate dehydrogenase (FDH) and mannitol dehydrogenase (MDH), for a cascade reaction producing d-mannitol. Coencapsulation of FDH and MDH resulted in an over 10-fold increase in d-mannitol production compared to the free mix of FDH and MDH, likely due to the enhanced local concentrations of FDH and MDH inside the PNC.

In response to the COVID-19 pandemic, different types of vaccines, such as inactive, live-attenuated, messenger RNA (mRNA), and protein subunit, have been developed against SARS-CoV-2. This has unintentionally created a unique scenario where heterologous prime-boost vaccination against a single virus has been administered to a large human population. Here, we aimed to analyze whether the immunization order of vaccine types influences the efficacy of heterologous prime-boost vaccination, especially mRNA and protein-based vaccines. We developed a new mRNA vaccine encoding the hemagglutinin (HA) glycoprotein of the influenza virus using the 3′-UTR and 5′-UTR of muscle cells (mRNA-HA) and tested its efficacy by heterologous immunization with an HA protein vaccine (protein-HA). The results demonstrated higher IgG2a levels and hemagglutination inhibition titers in the mRNA-HA priming/protein-HA boosting (R-P) regimen than those induced by reverse immunization (protein-HA priming/mRNA-HA boosting, P-R). After the viral challenge, the R-P group showed lower virus loads and less inflammation in the lungs than the P-R group did. Transcriptome analysis revealed that the heterologous prime-boost groups had differentially activated immune response pathways, according to the order of immunization. In summary, our results demonstrate that the sequence of vaccination is critical to direct desired immune responses. This study demonstrates the potential of a heterologous vaccination strategy using mRNA and protein vaccine platforms against viral infection.

Mercury complex of 4-(2-pyridylazo)resorcinol (PAR-2Hg(2+)), a halide-ion chemosensor, was prepared and its efficiency as a tool for high-throughput screening (HTS) of transition-metal-catalyzed coupling reactions was investigated. It showed a high selectivity for halide ions. When the PAR-2Hg(2+) complex was used in the Suzuki coupling reaction and C-H activated coupling reaction with aryl bromides, the quantitative and qualitative conversions of aryl halides were obtained from the reaction mixture color change.

No abstract

A paper-type sensor system was devised using an enzyme-loaded, artificial peroxidase-conjugated nanocarrier to maintain long-term stability with smartphone readout.

The development of artificial peroxidases has attracted great interest because of their applications in various fields such as the chemical industry and biosensing. In this study, 2,6-bis[(bis(2-pyridylmethyl)amino)-methyl]-4-methylphenol (H-bpmp) complexes with various transition metal ions have been investigated as artificial peroxidases. Among these metal complexes, the [Mn2(bpmp)]3+ complex showed the highest peroxidase-like activity as determined by a colorimetric assay using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and H2O2. The peroxidase-like activity was inhibited by pyrophosphate (PPi), which blocked the active site of the [Mn2(bpmp)]3+ complex. Based on this phenomenon, the ABTS/H2O2/[Mn2(bpmp)]3+ system could be applied for the detection of PPi, which could be achieved selectively by visual observation with a detection limit of 130 nM. Moreover, the addition of pyrophosphatase (PPase) to the [Mn2(bpmp)]3+ complex blocked by PPi resulted in the recovery of the peroxidase-like activity of the [Mn2(bpmp)]3+ complex due to the hydrolysis of PPi. Hence, the enzyme cascade reaction of the PPase and [Mn2(bpmp)]3+ complex allowed the real-time colorimetric assay of PPase.

No abstract