For the medical diagnosis of sepsis, it is crucial to differentiate infectious inflammation from noninfectious symptoms to prevent acute aggravation. Herein, a diagnosis for early stage sepsis was performed using LPC 16:0 and total phospholipids as small molecular biomarkers. The measurement of LPC 16:0 was conducted using a parylene matrix chip, which was developed to effectively detect small molecules in laser desorption/ionization mass spectrometry (LDI-MS).
View Article and Find Full Text PDFBackground: Hepatocellular carcinoma (HCC) stands as the sixth most prevalent cancer globally, presenting a substantial health challenge, particularly due to late-stage diagnoses that limit treatment effectiveness. Sorafenib, a multi-kinase inhibitor, is the primary chemotherapeutic agent for advanced HCC, but it only extends survival by 2-3 months. However, drug resistance remains a major clinical challenge, necessitating the exploration of new molecular mechanisms, including the role of microRNAs (miRNAs) in sorafenib resistance.
View Article and Find Full Text PDFFor the prevention of SARS-CoV-2 infection, four Fv-antibodies with binding affinity for the ACE2 receptor were screened from an Fv-antibody library. The screened Fv-antibodies were expressed as soluble proteins and estimated to have a high binding affinity, comparable to that between SARS-CoV-2 and the ACE2 receptor. The interaction between the Fv-antibodies and the ACE2 receptor was analyzed using docking simulation, and the significant binding affinity of the screened Fv-antibodies was attributed to the homology in amino acid sequence with the ACE2 receptor.
View Article and Find Full Text PDFIn this study, the vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was developed using ferritin complexes with the immunogenic sequences screened against the SARS-CoV-2 spike protein (SP) from the Fv-antibody library. The Fv-antibody library was prepared on the outer membrane of by the expression of the V region of immunoglobulin G (IgG) with a randomized complementarity-determining region 3 (CDR3). Four Fv-antibodies to the receptor-binding domain (RBD) were screened from the Fv-antibody library, which had a comparable binding constant () between SARS-CoV-2 SP and the angiotensin-converting enzyme 2 (ACE2) receptor.
View Article and Find Full Text PDFMigraine is known to be caused by calcitonin gene-related peptide (CGRP), prompting the need for quantitative analysis of CGRP for the clinical treatment of monoclonal antibodies targeting CGRP. Since CGRP is cleaved by proteolytic enzymes post-blood collection, rapid analysis methods are required. In this study, a one-step immunoassay for CGRP was developed using chemically mimicking peptides (mimotopes) with an analysis time of 32 min.
View Article and Find Full Text PDFACS Appl Mater Interfaces
November 2024
Parylene-like films obtained via the plasma decomposition of parylene precursors with functional groups (amino and formyl) are proposed as an alternative to those obtained via the thermal method. To analyze the chemical functional groups after plasma deposition, a surface analysis of the parylene films using the two different deposition methods was performed via Fourier transform infrared (FT-IR) and X-ray photoelectron spectroscopy (XPS). The FT-IR analysis revealed that the featured peaks of the chemical functional groups were maintained in the parylene-like films obtained via the plasma deposition method.
View Article and Find Full Text PDFThe emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants presents challenges in global efforts to transition from the pandemic to an endemic stage. The spike protein of the SARS-CoV-2 virus, which is pivotal for cell entry, exhibits significant mutations in its variants, potentially affecting infectivity and therapeutic efficacy. Recent findings indicate upregulation of the epidermal growth factor receptor (EGFR) pathway, a key target in cancer therapy, by the spike protein of SARS-CoV-2.
View Article and Find Full Text PDFFv-antibodies targeting the proprotein convertase (PPC) region of the SARS-CoV-2 spike protein (SP) were screened from an Fv-antibody library to inhibit SARS-CoV-2 infection. Two selected Fv-antibodies were expressed as soluble recombinant proteins, and their binding affinities were assessed using a surface plasmon resonance biosensor. The binding regions of these Fv-antibodies corresponded to the cleavage sites of furin (S1/S2) and transmembrane serine protease 2 (TMPRSS2, S2').
View Article and Find Full Text PDFInhibitors of the epithermal growth factor receptor (EGFR) were screened from an autodisplayed Fv-antibody library using an anti-EGF antibody. The Fv-antibody library was expressed on the outer membrane of , which corresponds to the heavy chain V region of immunoglobulin G. The library was constructed by randomizing the CDR3 region of expressed V regions (11 amino acid residues) by site-directed mutagenesis.
View Article and Find Full Text PDFIn this study, a one-step immunoassay for porcine epidemic diarrhea virus (PEDV) based on Fv-antibodies and switching peptides was developed, and the assay results of PEDV were obtained by just mixing samples without any further reaction or washing steps. The Fv-antibodies with binding affinity to the spike protein of PEDV were screened from the Fv-antibody library using the receptor-binding domain (RBD) of the spike protein as a screening probe. Screened Fv-antibodies with binding affinities to the RBD antigen were expressed, and the binding constants () were calculated to be 83-142 nM.
View Article and Find Full Text PDFMonocarboxylate transporter-1 (MCT-1) inhibitors were screened from the Fv-antibody library, which contained complementary determining region 3 with randomized amino acid sequences (11 residues) through site-directed mutagenesis. Fv-antibodies against MCT-1 were screened from the autodisplayed Fv-antibody library. Two clones were screened, and the binding affinity (K) against MCT-1 was estimated using flow cytometry.
View Article and Find Full Text PDFOuter membrane vesicle-functionalized nanoparticles (OMV-NPs) have attracted significant interest, especially regarding drug delivery applications and vaccines. Here, we report on novel OMV-NPs by applying bioorthogonal click reaction for encapsulating gold nanoparticles (NPs) within outer membrane vesicles (OMVs) by covalent coupling. For this purpose, outer membrane protein A (OmpA), abundant in large numbers (due to 100,000 copies/cell [1]) in OMVs, was modified via the incorporation of the unnatural amino acid p-azidophenylalanine.
View Article and Find Full Text PDFIn several fields, the process of fusing multiple two-dimensional (2D) closed lines is an important step. For instance, this is fundamental in histology and oncology in general. The treatment of a tumor consists of numerous steps and activities.
View Article and Find Full Text PDFHepatocellular carcinoma (HCC) is the fifth leading cause of cancer-related mortality worldwide. Programmed cell death ligand-1 (PD-L1) is an immune checkpoint protein that binds to programmed cell death-1 (PD-1), which is expressed in activated T cells and other immune cells and has been employed in cancer therapy, including HCC. Recently, PD-L1 overexpression has been documented in treatment-resistant cancer cells.
View Article and Find Full Text PDFACS Pharmacol Transl Sci
January 2024
Serotonin-like mimotopes were screened from the Fv-antibody library to be used as inhibitors against monoamine oxidase A (MAO-A). The Fv-antibody [corresponding to the V region of immunoglobulin G (IgG)] consists of three complementarity-determining regions and four frame regions. The Fv-antibody library was prepared by site-directed mutagenesis of CDR3, which consists of 11 amino acid residues.
View Article and Find Full Text PDFACS Pharmacol Transl Sci
December 2023
Pancreatic ribonuclease A (RNase A) inhibitors were screened from an autodisplayed Fv-antibody library, which was prepared by randomizing amino acid sequences of the third complementary-determining region (CDR3) within the heavy chain variable region (V region) of immunoglobulin G (called "Fv-antibody" comprising three CDRs and four frame regions (FRs)) through site-directed mutagenesis. The library was autodisplayed on the outer membrane of . Target Fv-variants (clones) with specific binding affinity for RNase A were screened using fluorescein-labeled RNase A and flow cytometry.
View Article and Find Full Text PDFA defect-passivated photosensor based on cesium lead bromide (CsPbBr) perovskite quantum dots (QD) was fabricated using parylene films, and the photosensor was applied for the microbial detection. The CsPbBr perovskite QDs were synthesized to be homogeneous in size under thermodynamic control, and the perovskite QD-based photosensor was fabricated using MoS flakes as the electron transfer layer. In this work, a parylene film with functional groups was deposited on a photosensor for physical protection (waterproof) and defect (halide vacancy) passivation of the perovskite QD.
View Article and Find Full Text PDFThe Fv-antibodies were correponded to V region of immunoglobulin G, which were composed of three complementarity determining regions (CDRs) for the specific binding of antigens. In this work, the Fv-antibodies against SARS-CoV-2 spike protein (SP) were screened from an autodisplayed Fv-antibody library which was expressed on E. coli outer membrane, and the receptor binding domain (RBD) of SP was used as a screening probe.
View Article and Find Full Text PDFElectrochemical analysis of total phospholipids was performed for the diagnosis of sepsis. The influence of electrode materials on the analysis of the chromogenic substrate was analyzed using Au, graphite, and pyrolyzed carbon electrodes. The total phospholipid analysis based on electrochemical analysis with pyrolyzed carbon was used for diagnosis of sepsis using sera from healthy volunteers, systemic inflammatory response syndrome (SIRS), and severe sepsis patients.
View Article and Find Full Text PDFWearable devices that can mechanically conform to human skin are a necessity for reliable monitoring and decoding of biomechanical activities through skin. Most inorganic piezoelectrics, however, lack deformability and damage tolerance, impeding stable motion monitoring. Here, we present an air-permeable fabric-based ZnO nanogenerator with mechanical adaptivity to diverse deformations for wearable piezoelectric sensors, collecting biomechanical health data.
View Article and Find Full Text PDFVariants in SLC26A4 (pendrin) are the most common reasons for genetic hearing loss and vestibular dysfunction in East Asians. In patients with Pendred syndrome and DFNB4 (autosomal recessive type of genetic hearing loss 4), caused by variants in SLC26A4, the hearing function is residual at birth and deteriorates over several years, with no curative treatment for these disorders. In the present study, we revealed that a novel small molecule restores the expression and function of mutant pendrin.
View Article and Find Full Text PDFIn this study, the influence of microenvironments on antibody production of hybridoma cells was analyzed using six types of functionalized parylene films, parylene-N and parylene-C (before and after UV radiation), parylene-AM, and parylene-H, and using polystyrene as a negative control. Hybridoma cells were cultured on modified parylene films that produced a monoclonal antibody against the well-known fungal toxin ochratoxin-A. Surface properties were analyzed for each parylene film, such as roughness, chemical functional groups, and hydrophilicity.
View Article and Find Full Text PDFThe use of phages-a natural predator of bacteria-has emerged as a therapeutic strategy for treating multidrug-resistant bacterial infections; thus, the isolation and detection of phages from the environment is crucial for advancing phage therapy. Herein, for the first time, we propose a nanoplasmonic-based biodetection platform for phages that utilizes bacterial outer membranes (OMs) as a biorecognition element. Conventional biosensors based on phage-bacteria interactions encounter multiple challenges due to the bacteriolytic phages and potentially toxic bacteria, resulting in instability and risk in the measurement.
View Article and Find Full Text PDFIn this work, we investigated the effect of hole transporting poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) interfacing with Mn-doped CdS/ZnS quantum dots (QDs) deposited on an indium tin oxide (ITO) substrate on the photoemission of upconverted hot electrons under weak continuous wave photoexcitation in a vacuum. Among the various factors that can influence the photoemission of the upconverted hot electrons, we studied the role of PEDOT:PSS in facilitating the hole transfer from QDs and altering the energy of photoemitted hot electrons. Compared to hot electrons emitted from QDs deposited directly on the ITO substrate, the addition of the PEDOT:PSS layer between the QD and ITO layers increased the energy of the photoemitted hot electrons.
View Article and Find Full Text PDFThe detection of severe acute respiratory syndrome coronavirus (SARS-CoV-1) was demonstrated using screened Fv-antibodies for SPR biosensor and impedance spectrometry. The Fv-antibody library was first prepared on the outer membrane of E. coli using autodisplay technology and the Fv-variants (clones) with a specific affinity toward the SARS-CoV-1 spike protein (SP) were screened using magnetic beads immobilized with the SP.
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