Facile biotic/abiotic sandwich detection system for the highly sensitive detection of human serum albumin and glycated albumin.

Quantifying glycated albumin (GA) levels in the blood is crucial for diagnosing diabetes because they strongly correlate with blood glucose concentration. In this study, a biotic/abiotic sandwich assay was developed for the facile, rapid, and susceptible detection of human serum albumin (HSA) and GA. The proposed sandwich detection system was assembled using a combination of two synthetic polymer receptors and natural antibodies.

A rapid abiotic/biotic hybrid sandwich detection for trace pork adulteration in halal meat extract.

In this study, we prepared molecularly imprinted polymer nanogels with good affinity for the Fc domain of immunoglobulin G (IgG) using 4-(2-methacrylamidoethylaminomethyl) phenylboronic acid as a modifiable functional monomer for post-imprinting in-cavity modification of a fluorescent dye (F-Fc-MIP-NGs). A novel nanogel-based biotic/abiotic hybrid sandwich detection system for porcine serum albumin (PSA) was developed using F-Fc-MIP-NGs as an alternative to a secondary antibody for fluorescence detection and another molecularly imprinted polymer nanogel capable of recognizing PSA (PSA-MIP-NGs) as a capturing artificial antibody, along with a natural antibody toward PSA (Anti-PSA) that was used as a primary antibody. After incubation of PSA and Anti-PSA with F-Fc-MIP-NGs, the PSA/Anti-PSA/F-Fc-MIP-NGs complex was captured by immobilized PSA-MIP-NGs for fluorescence measurements.

A molecularly imprinted nanocavity with transformable domains that fluorescently indicate the presence of antibiotics in meat extract samples.

In this study, we aimed to create synthetic polymer receptors with the fluorescence signalling ability, using molecular imprinting, precisely designed template molecules, and site-specific post-imprinting modifications, which can mimic conjugated proteins and are capable of specific molecular recognition, and wherein successful binding can be indicated by a change in fluorescence. A molecularly imprinted APO-type nanocavity with a reconstructable domain was prepared by co-polymerisation of a template molecule containing cephalexin conjugated to polymerisable groups a Schiff base, a disulphide bond, and a cross-linker, followed by hydrolysis of the Schiff base and a disulphide exchange reaction. Fluorescence-based indication of binding was devised by the Schiff base formation reaction with 4-formylsalicylic acid, and the interacting site was introduced a disulphide exchange reaction with 4-mercaptobenzoic acid, yielding a multifunctional mature (HOLO)-type molecularly imprinted nanocavity.

Biocompatible polymer-modified gold nanocomposites of different shapes as radiation sensitizers.

Radiation therapy is a powerful approach for cancer treatment due to its low invasiveness. The development of radiation sensitizers is of great importance as they assist in providing radiation therapy at a low dose. In this study, poly(2-methacryloyloxyethyl phosphorylcholine) (PMPC)-modified gold nanocomposites of different shapes were created using the grafting-to approach to serve as a novel radiation sensitizer with high cellular uptake.

Fc Domain-Imprinted Stealth Nanogels Capable of Orientational Control of Immunoglobulin G Adsorbed In Vivo.

Regulation of nanomaterial-cell interaction is an important requisite for a variety of biomedical applications such as drug delivery systems and theranostics. Here, we demonstrate the regulation of nanomaterial-cell interaction using the oriented adsorption of intrinsic immunoglobulin G (IgG) on molecularly imprinted polymer nanogels (MIP-NGs) capable of recognizing the fragment crystallizable (Fc) domain of IgG. The unique domain recognition property resulted in the suppression of the immune response in Fc domain receptor-possessing macrophages and natural killer cells due to the regulation of protein corona based on the oriented adsorption of IgG.

Protein-imprinted polymer films prepared via cavity-selective multi-step post-imprinting modifications for highly selective protein recognition.

The use of molecularly imprinted polymers (MIPs) for achieving synthetic receptors capable of selective molecular recognition is promising; however, these polymers exhibit low selectivity derived from the heterogeneity of their created, imprinted cavities. To achieve highly selective protein recognition, we herein report the cavity-selective, multi-step, post-imprinting modification of MIPs. An MIP film for lysozyme was prepared by the copolymerization of {[2-(2-methacrylamido)ethyldithio]ethylcarbamoyl}methoxy acetic acid, a functional monomer possessing a modifiable disulfide bond, with acrylamide and N,N'-methylenebisacrylamide in the presence of lysozyme.

Molecularly imprinted polymer nanogel-based fluorescence sensing of pork contamination in halal meat extracts.

Pork contamination is a serious concern for the global halal food market because many manufacturers commonly use pork instead of beef to reduce production costs. In this study, a highly sensitive fluorescent molecularly imprinted polymer nanogel (F-MIP-NG)-based sensor was developed for rapid porcine serum albumin (PSA) detection to investigate pork contamination in halal meat extracts. F-MIP-NGs were prepared via molecular imprinting and conjugation with ATTO 647N as the fluorescent reporter molecule for the post-imprinting modification (PIM) and then immobilized on gold-coated sensor chips.

Molecularly Imprinted Nanogels Possessing Dansylamide Interaction Sites for Controlling Protein Corona In Situ by Cloaking Intrinsic Human Serum Albumin.

Nanomaterials have become increasingly promising for biomedical applications owing to their specific biological characteristics. As drug delivery vehicles, nanomaterials have to circulate in the bloodstream to deliver the encapsulated components to the target tissues. Protein corona regulation is one of the promising approaches that gives stealth capability to avoid immune response.

Signalling molecular recognition nanocavities with multiple functional groups prepared by molecular imprinting and sequential post-imprinting modifications for prostate cancer biomarker glycoprotein detection.

Fluorescent-signalling molecularly-imprinted nanocavities possessing orthogonal dual interaction sites for the detection of prostate cancer biomarker glycoprotein were constructed through molecular imprinting and sequential multistep post-imprinting modifications (PIMs) using a newly designed multi-functionalised PIM reagent (PIR). The PIR, possessing an interaction site and dual reaction sites for PIMs, enabled us to introduce multiple functions including interaction sites and fluorescent reporter groups in a single PIM site, leading to the sensitive fluorescent detection of target glycoproteins with a high signal-to-noise ratio. Prostate specific antigen (PSA), used as a biomarker for prostate-related diseases, was selected as a target glycoprotein.

Molecularly Imprinted Nanogels Capable of Porcine Serum Albumin Detection in Raw Meat Extract for Halal Food Control.

Accurate, simple, and valuable analytical methods for detection of food contamination are rapidly expanding to evaluate the validity of food product quality because of ethnic considerations and food safety. Herein molecularly imprinted nanogels (MIP-NGs), capable of porcine serum albumin (PSA) recognition, were prepared as artificial molecular recognition elements. The MIP-NGs were immobilized on a quartz crystal microbalance (QCM) sensor for detection of pork contamination in real beef extract samples.

Antibody-Conjugated Signaling Nanocavities Fabricated by Dynamic Molding for Detecting Cancers Using Small Extracellular Vesicle Markers from Tears.

Small extracellular vesicles (sEVs) are reliable biomarkers for early cancer detection; however, conventional detection methods such as immune-based assays and microRNA analyses are not very sensitive and require sample pretreatments and long analysis time. Here, we developed a molecular imprinting-based dynamic molding approach to fabricate antibody-conjugated signaling nanocavities capable of size recognition. This enabled the establishment of an easy-to-use, rapid, sensitive, pretreatment-free, and noninvasive sEV detection platform for efficient sEV detection-based cancer diagnosis.

High Mannose Binding Lectin (PFL) from Down-Regulates Cancer-Associated Integrins and Immune Checkpoint Ligand B7-H4.

lectin (PFL), which belongs to the high mannose (HM)-binding OAAH ( agglutinin homologue) lectin family, induces cancer cell death. However, the detailed mechanisms underlying this process have not yet been elucidated. We found that PFL decreased various integrins as well as EGFR in cancer cells by promoting internalization and autophagic degradation of these molecules, subsequently inducing caspase-8 dependent cell apoptosis.

Site-specific post-imprinting modification of molecularly imprinted polymer nanocavities with a modifiable functional monomer for prostate cancer biomarker recognition.

Recognition of glycans of glycoproteins using biotic materials such as antibodies is challenging due to lack of antigenicity. Polymeric materials suitable for the molecular recognition of glycoproteins have attracted considerable attention. In this study, we aimed to develop abiotic molecular materials for the recognition of prostate-specific antigen (PSA), a known biomarker for prostate cancer.

A Pretreatment-Free, Polymer-Based Platform Prepared by Molecular Imprinting and Post-Imprinting Modifications for Sensing Intact Exosomes.

Exosomes are small (30-100 nm) membrane vesicles that serve as regulatory agents for intercellular communication in cancers. Currently, exosomes are detected by immuno-based assays with appropriate pretreatments like ultracentrifugation and are time consuming (>12 h). We present a novel pretreatment-free fluorescence-based sensing platform for intact exosomes, wherein exchangeable antibodies and fluorescent reporter molecules were aligned inside exosome-binding cavities.

Orientationally Fabricated Zwitterionic Molecularly Imprinted Nanocavities for Highly Sensitive Glycoprotein Recognition.

Glycoprotein recognition has recently gained a lot of attention, since glycoproteins play important roles in a diverse range of biological processes. Robustly synthesized glycoprotein receptors, such as molecularly imprinted polymers (MIPs), which can be easily and sustainably handled, are highly attractive as antibody substitutes because of the difficulty in obtaining high-affinity antibodies specific for carbohydrate-containing antigens. Herein, molecularly imprinted nanocavities for glycoproteins have been fabricated via a bottom-up molecular imprinting approach using surface-initiated atom transfer radical polymerization (SI-ATRP).

Beyond natural antibodies - a new generation of synthetic antibodies created by post-imprinting modification of molecularly imprinted polymers.

The molecular imprinting technology yields artificial materials capable of antibody-like molecular recognition. Molecularly imprinted materials are attractive because procedures for their preparation and use are comparatively simple. The number of research reports concerning molecularly imprinted polymers (MIPs) have been increasing yearly, attracting a great deal of interest in various fields.

Oriented, molecularly imprinted cavities with dual binding sites for highly sensitive and selective recognition of cortisol.

Novel, molecularly imprinted polymers (MIPs) were developed for the highly sensitive and selective recognition of the stress marker cortisol. Oriented, homogeneous cavities with two binding sites for cortisol were fabricated by surface-initiated atom transfer radical polymerization, using a cortisol motif template molecule (TM1) which consists of a polymerizable moiety attached at the 3-carbonyl group of cortisol via an oxime linkage and an adamantane carboxylate moiety coupled with the 21-hydroxyl group. TM1 was orientationally immobilized on a βcyclodextrin (β-CD)-grafted gold-coated sensor chip by inclusion of the adamantane moiety of TM1, followed by copolymerization of a hydrophilic comonomer, 2-methacryloyloxyethyl phosphorylcholine, with or without a cross-linker, -methylenebisacrylamide.

Regulation of protein-binding activities of molecularly imprinted polymers via post-imprinting modifications to exchange functional groups within the imprinted cavity.

We prepared lysozyme-imprinted polymers bearing modifiable sites within the imprinted cavity to introduce various functional groups via post-imprinting modifications. For this purpose, ({[2-(2-methacrylamido)-ethyldithio]-ethylcarbamoyl}-methoxy)acetic acid (MDTA), which has a carboxy group to interact with the target protein, lysozyme, and a disulfide linkage for post-imprinting modifications, was used as a functional monomer. A lysozyme-imprinted polymer film was prepared by copolymerization of MDTA with a cross-linker, N,N'-methylenebisacrylamide, in the presence of lysozyme.

Regioselective Molecularly Imprinted Reaction Field for [4 + 4] Photocyclodimerization of 2-Anthracenecarboxylic Acid.

Molecularly imprinted cavities have functioned as a regioselective reaction field for the [4 + 4] photocyclodimerization of 2-anthracenecarboxylic acid (2-AC). Molecularly imprinted polymers were prepared by precipitation polymerization of N-methacryloyl-4-aminobenzamidine as a functional monomer to form a complex with template 2-AC and ethylene glycol dimethacrylate as a crosslinking monomer. The 2-AC-imprinted cavities thus constructed preferentially bound 2-AC with an affinity greater than that toward structurally related 9-anthracenecarboxylic acid, 2-aminoanthracene, and unsubstituted anthracene.

Fluorescence signaling molecularly imprinted polymers for antibiotics prepared via site-directed post-imprinting introduction of plural fluorescent reporters within the recognition cavity.

An antibiotic-imprinted cavity with two different fluorescent dyes was prepared by molecular imprinting and subsequent post-imprinting modifications (PIMs), for the readout of a specific binding event as a fluorescence signal. The fluorescent dyes were site-specifically introduced into the cavity using an orthogonal reversible bonding reaction, Schiff base formation, and a disulfide exchange reaction. The template molecule, comprising cephalexin connected to a Schiff base monomer and a disulfide monomer, was copolymerized with a crosslinker.

A Programmable Signaling Molecular Recognition Nanocavity Prepared by Molecular Imprinting and Post-Imprinting Modifications.

Inspired by biosystems, a process is proposed for preparing next-generation artificial polymer receptors with molecular recognition abilities capable of programmable site-directed modification following construction of nanocavities to provide multi-functionality. The proposed strategy involves strictly regulated multi-step chemical modifications: 1) fabrication of scaffolds by molecular imprinting for use as molecular recognition fields possessing reactive sites for further modifications at pre-determined positions, and 2) conjugation of appropriate functional groups with the reactive sites by post-imprinting modifications to develop programmed functionalizations designed prior to polymerization, allowing independent introduction of multiple functional groups. The proposed strategy holds promise as a reliable, affordable, and versatile approach, facilitating the emergence of polymer-based artificial antibodies bearing desirable functions that are beyond those of natural antibodies.

Post-imprinting and In-Cavity Functionalization.

Molecularly imprinted polymers (MIPs) are artificial materials capable of molecular recognition for target molecules. Currently MIPs have been prepared without further modification after polymerization, and used for predetermined single purposes. Post-imprinting modifications (PIMs) presented here can provide site-specific modifications within the molecularly imprinted binding cavities after polymerization, enabling MIPs to become more complex functional materials as were the cases of naturally occurring conjugated proteins.

Preparation of molecularly imprinted polymers for the recognition of proteins via the generation of peptide-fragment binding sites by semi-covalent imprinting and enzymatic digestion.

Molecularly imprinted polymers bearing peptide fragment-based binding sites within the protein-imprinted cavities were prepared by copolymerization of the acrylated protein with 6-monoacryloyl-trehalose and 6,6'-diacryloyl-trehalose as a hydrophilic comonomer and a crosslinker respectively, followed by enzymatic decomposition of the grafted protein into the polymer matrix with pepsin, resulting in the creation of peptide fragment-based protein-binding sites.