Cancer-derived circulating exosomes or nanoscale extracellular vesicles are emerging biomarkers for disease detection and treatment because of their cell-specific constituents and unique ...intercellular pathways. For efficient exosome isolation from bio-fluids, the design of high-affinity nanointerfaces is of great importance in the development of miniaturized systems for the collection of exosomes. Herein, we report peptide-functionalized nanowires as a biorecognition interface for the capture and release of cancer-derived exosomes within a microfluidic channel. Based on the amino-acid sequence of EWI-2 protein, a partial peptide that bound to the CD9 exosome marker and thus targeted cancer exosomes was screened. Linkage of the exosome-targeting peptide with a ZnO-binding sequence allowed one-step and reagent-free peptide modification of the ZnO nanowire array. As a result of peptide functionalization, the exosome-capturing ability of ZnO nanowires was significantly improved. Furthermore, the captured exosomes could be subsequently released from the nanowires under a neutral salt condition for downstream applications. This engineered surface that enhances the nanowires' efficiency in selective and controllable collection of cancer-derived exosomes provides an alternative foundation for developing microfluidic platforms for exosome-based diagnostics and therapeutics.
Membrane curvature-sensing (MCS) proteins recognize and regulate the morphologies of biological membranes. As these proteins lack characteristic sequence motifs in their primary structure, they are ...not instantly recognizable by genomic databases. Overcoming this technological challenge toward the agile identification of new proteins can promote the elucidation of membrane morphological regulation. Here, for the selective identification of MCS proteins, comparative proteomic analysis was performed using different sizes of the spherical supported lipid bilayer (SSLB), which consists of spherical SiO2 particles covered with a lipid bilayer. Because of the presence of SiO2 core, the curvature of the surrounding membrane is well-controlled and stable even on a micron scale. To prove this concept, known membrane curvature-sensing protein domains, Bin/Amphiphysin/Rvs (BAR) and Epsin N-terminal homology (ENTH), were evaluated by performing a binding assay using SSLBs, and the preferential binding to the highly curved membrane was confirmed. Peripheral membrane proteins obtained from normal human dermal fibroblast (NHDF) and human breast cancer (MDA-MB-231) cells were used in shotgun proteomic analysis, and 786 and 949 proteins were identified from SSLBs as lipid membrane binders, respectively. Statistical quantitative analyses of proteins detected from each SSLB with a different size revealed 118 candidate proteins, including 23 proteins unique to MDA-MB-231 cells, as membrane curvature sensors, including some previously reported curvature sensors. Functional clustering analysis based on the KEGG orthology database revealed that the protein-binding property to specific high or low membrane curvature correlated with their functions. Further investigation of candidate proteins will lead to the identification of new MCS proteins as well as cancer biomarkers.
A CD9-binding peptide (RSHRLRLH), screened from EWI-2, was characterized, and its effect on cellular migration and invasion was evaluated. As CD9 protein is overexpressed in cancer cells and plays an ...important role in cellular migration, the CD9-binding peptide preferentially inhibited the migration of cancer cells. Unlike conventional antiproliferative drugs, this CD9-binding peptide is promising as a novel precision antimigratory agent for cancer therapeutics.
A CD9-binding peptide (RSHRLRLH), screened from EWI-2, was characterized, and its inhibition effect on cancer-cell migration and invasion was demonstrated.
Dipeptidyl peptidase IV (DPP-IV) has become an important target in the prevention and treatment of diabetes. Although many DPP-IV inhibitory peptides have been identified by a general approach ...involving the repeated fractionation of food protein hydrolysates, the obtained results have been dependent on the content of each peptide and fractionation conditions. In the present study, a peptide array that provides comprehensive assays of peptide sequences was used to identify novel DPP-IV inhibitory peptides derived from bovine milk proteins; these peptides were then compared with those identified using the general approach. While the general approach identified only known peptides that were abundant in the hydrolysate, the peptide array-based approach identified 10 novel DPP-IV inhibitory peptides, all of which had proline at the second residue from the N-terminus. The proper or combined use of these two approaches, which have different advantages, will enable the efficient development of novel bioactive foods and drugs.
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•Peptide array can be used to screen DPP-IV inhibitory peptides from food proteins.•Ten novel DPP-IV inhibitory peptides were identified.•All the novel peptides contained a proline at the second residue from the N-terminus.
Cancer immunotherapy is an emerging therapeutic strategy for cancer treatment. Most of the immunotherapeutics approved by the FDA regulate the innate immune system and associated immune cell ...activity, with immune check inhibitors in particular having transformed the field of cancer immunotherapy due to their significant clinical potential. However, previously reported immunotherapeutics have exhibited undesirable side effects, including autoimmune toxicity and inflammation. Controlling these deleterious responses and designing therapeutics that can precisely target specific regions are thus crucial to improving the efficacy of cancer immunotherapies. Recent studies have reported that cancer cells employ glycan−immune checkpoint interactions to modulate immune cell activity. Thus, the recognition of cancer glycan moieties such as sialoglycans may improve the anticancer activity of immune cells. In this review, we discuss recent advances in cancer immunotherapies involving glycans and glycan-targeting technologies based on nanomaterial-assisted local delivery systems.
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•Cancer cells employ glycan−immune checkpoint interactions.•The recognition of cancer glycan moieties may improve the anticancer activity.•Glycans and glycan-targeting technologies may advance cancer immunotherapies.
•The TNT recognition peptide candidates were rationally designed and identified through the complementarity determining region (CDR) of anti-TNT monoclonal antibody. Surface Plasmon resonance (SPR) ...sensor functionalized with peptide candidates was utilized to screen the TNT binding peptide and characterize its sensitivity and selectivity to TNT explosive.
In this study, 2,4,6-trinitrotoluene (TNT) binding peptide was synthesized and screened for TNT specific detection using surface plasmon resonance (SPR) sensor. The TNT binding peptide was rational design and synthesized through amino acid sequence from complementarity determining region (CDR) in the anti-TNT monoclonal antibody, which was produced from hybridoma cell using TNP-KLH conjugate as antigen. Three TNT binding peptide sequences were obtained from the heavy chain of CDR1 named TNTHCDR1, TNTHCDR2 from CDR2 and TNTHCDR3 from CDR3 of anti-TNT antibody. Screening process of three candidate peptides were carried out by using the SPR sensor with direct determination, which the peptide was directly immobilized on the sensor chip CM7 surface through amine coupling reaction. The results demonstrated that peptide TNTHCDR3 was determined as TNT binding peptide and no non-specific binding was observed. The selectivity of TNT binding peptide TNTHCDR3 was also testified by six kinds of TNT analogues. The results indicated the specific binding between TNT and peptide TNTHCDR3.
With increased awareness among consumers regarding food safety and security, food allergen control has become an indispensable requirement in the food industry. Although several methods for detecting ...allergens in food products are available, highly sensitive techniques are required. In this study, we developed a technique named as peptide array-based inhibition enzyme-linked immunosorbent assay (ELISA), Pep-iEIA, for evaluating antigenicity and detecting cow's milk antigen in infant formula products, using a peptide array consisting of a series of overlapping peptides found in allergenic milk proteins. Pep-iEIA was used to examine five cow's milk-based infant formulas with different degrees of hydrolyzation, and the assay offered both more sensitive detection and detailed analysis of remaining antigenic peptides in allergen compared to conventional ELISA. The antigenicity level of the allergenic peptides identified using Pep-iEIA was confirmed by surface plasmon resonance assay. We believe that Pep-iEIA will be highly useful for antigenicity evaluation of dairy products consumed by infants and patients with cow's milk allergy.
Rapid diagnosis of flu before symptom onsets can revolutionize our health through diminishing a risk for serious complication as well as preventing infectious disease outbreak. Sensor sensitivity and ...selectivity are key to accomplish this goal as the number of virus is quite small at the early stage of infection. Here we report on label-free electrical diagnostics of influenza based on nanopore analytics that distinguishes individual virions by their distinct physical features. We accomplish selective resistive-pulse sensing of single flu virus having negative surface charges in a physiological media by exploiting electroosmotic flow to filter contaminants at the Si
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pore orifice. We demonstrate identifications of allotypes with 68% accuracy at the single-virus level via pattern classifications of the ionic current signatures. We also show that this discriminability becomes >95% under a binomial distribution theorem by ensembling the pulse data of >20 virions. This simple mechanism is versatile for point-of-care tests of a wide range of flu types.
•Tritryptophan peptides self-assembled on a pyrolytic graphite/graphene surface.•The peptide interface did not alter electrochemical properties of pyrolytic graphite.•Redox signals of potassium ...ferricyanide were maintained.•Bi-functional peptides induced cell adhesion of anchorage-dependent cells.
Designing cell-electrode biointerfaces is important for electrochemical monitoring of cell functions and activities in their microenvironments. Herein, peptide monolayer was constructed on the pyrolytic graphite surface by self-assembling of tritryptophane, and the bi-functionalized peptide conjugating cell adhesion peptide, GRGDS (Gly-Arg-Gly-Asp-Ser) induced adhesion of anchorage-dependent cells. The peptide modification did not alter the electrochemical properties of pyrolytic graphite, and the redox signals of potassium ferricyanide were maintained. These observations indicate that non-covalent functionalization of graphite/graphene using bi-functional peptides would be effective for electrochemical measurement of cell activities.
In this study, we developed a surface plasmon resonance (SPR) sensor chip based on 2,4,6-trinitrotoluene (TNT) recognition peptide-modified single-walled carbon nanotubes (SWCNTs). The carboxylic ...acid-functionalized SWCNTs were immobilized on a 3-aminopropyltriethoxysilane (APTES)-modified SPR Au chip surface. Through π-stacking between the aromatic amino acids and SWCNTs, the TNT recognition peptide TNTHCDR3 was immobilized onto the surface of the SWCNTs. The peptide⁻SWCNTs-modified sensor surface was confirmed and evaluated by atomic force microscope (AFM) observation. The peptide⁻SWCNTs hybrid SPR sensor chip exhibited enhanced sensitivity with a limit of detection (LOD) of 772 ppb and highly selective detection compared with commercialized carboxymethylated dextran matrix sensor chips.
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