Construction and evaluation of AuNPs enhanced electrochemical immunosensors with [Fe(CN)6]3-/4- and PPy probe for highly sensitive detection of human chorionic gonadotropin.

Human chorionic gonadotropin (HCG), a vital protein for pregnancy determination and a marker for trophoblastic diseases, finds application in monitoring early pregnancy and ectopic pregnancy. This study presents an innovative approach employing electrochemical immunosensors for enhanced HCG detection, utilizing Anti-HCG antibodies and gold nanoparticles (AuNPs) in the sensor platform. Two sensor configurations were optimized: BSA/Anti-HCG/c-AuNPs/MEL/e-AuNPs/SPCE with [Fe(CN)6]3-/4- as a redox probe (1) and BSA/Anti-HCG/PPy/e-AuNPs/SPCE using polypyrrole (PPy) as a redox probe (2). The first sensor offers linear correlation in the 0.10-500.00 pg∙mL-1 HCG range, with a limit of detection (LOD) of 0.06 pg∙mL-1, sensitivity of 32.25 µA∙pg-1∙mL∙cm-2, RSD <2.47 %, and a recovery rate of 101.03-104.81 %. The second sensor widens the HCG detection range (40.00 fg∙mL-1-5.00 pg∙mL-1) with a LOD of 16.53 fg∙mL-1, ensuring precision (RSD <1.04 %) and a recovery range of 94.61-106.07 % in serum samples. These electrochemical immunosensors have transformative potential in biomarker detection, offering enhanced sensitivity, selectivity, and stability for advanced healthcare diagnostics.

Effects of amyloid-ß-mimicking peptide hydrogel matrix on neuronal progenitor cell phenotype.

Self-assembling peptide-based hydrogels have become a highly attractive scaffold for three-dimensional (3D) in vitro disease modeling as they provide a way to create tunable matrices that can resemble the extracellular matrix (ECM) of various microenvironments. Alzheimer's disease (AD) is an exceptionally complex neurodegenerative condition; however, our understanding has advanced due to the transition from two-dimensional (2D) to 3D in vitro modeling. Nonetheless, there is a current gap in knowledge regarding the role of amyloid structures, and previously developed models found long-term difficulty in creating an appropriate model involving the ECM and amyloid aggregates. In this report, we propose a multi-component self-assembling peptide-based hydrogel scaffold to mimic the amyloid-beta (ß) containing microenvironment. Characterization of the amyloid-ß-mimicking hydrogel (Col-HAMA-FF) reveals the formation of ß-sheet structures as a result of the self-assembling properties of phenylalanine (Phe, F) through π-π stacking of the residues, thus mimicking the amyloid-ß protein nanostructures. We investigated the effect of the amyloid-ß-mimicking microenvironment on healthy neuronal progenitor cells (NPCs) compared to a natural-mimicking matrix (Col-HAMA). Our results demonstrated higher levels of neuroinflammation and apoptosis markers when NPCs were cultured in the amyloid-like matrix compared to a natural brain matrix. Here, we provided insights into the impact of amyloid-like structures on NPC phenotypes and behaviors. This foundational work, before progressing to more complex plaque models, provides a promising scaffold for future investigations on AD mechanisms and drug testing. STATEMENT OF SIGNIFICANCE: In this study, we engineered two multi-component hydrogels: one to mimic the natural extracellular matrix (ECM) of the brain and one to resemble an amyloid-like microenvironment using a self-assembling peptide hydrogel. The self-assembling peptide mimics ß-amyloid fibrils seen in amyloid-ß protein aggregates. We report on the culture of neuronal progenitor cells within the amyloid-mimicking ECM scaffold to study the impact through marker expressions related to inflammation and DNA damage. This foundational work, before progressing to more complex plaque models, offers a promising scaffold for future investigations on AD mechanisms and drug testing.

Divalent metal ion modulation of a simple peptide-based hydrogel: self-assembly and viscoelastic properties.

Peptide self-assembly has been highly studied to understand the pathways in forming higher order structures along with the development and application of resulting hydrogel materials. Driven by noncovalent interactions, peptide hydrogels are stimuli-responsive to any addition to its gelling conditions. Here, a Phe-His based peptide, C14-FH(Trt)-OH, was synthesized and characterized with 1H NMR, FT-IR, MS, UV-vis spectroscopies and elemental analysis. Based on SEM imaging, the dipeptide conjugate was capable of forming a nanofibrous, interconnected network encapsulating buffer to produce a supramolecular hydrogel. Through the addition of Zn2+ and Cu2+, there is a clear change in the self-assembled nanostructures characterized through SEM. With this effect on self-assembly follows a change in the viscoelastic properties of the material, as determined through rheological frequency sweeps, with 2 and 3 orders of magnitude decreases in the elastic modulus G' in the presence of Zn2+ and Cu2+ respectively. This highlights the tunability of soft material properties with peptide design and self-assembly, through metal ions and Nδ-directed coordination.

Electrochemical immuno-biosensors for the detection of the tumor marker alpha-fetoprotein: A review.

Alpha-fetoprotein (AFP) is a glycoprotein that has many important physiological functions, including transportation, immunosuppression, and induction of apoptosis by T lymphocytes. AFP is closely related to the development of hepatocellular carcinoma and many kinds of tumors, all of which can show high concentrations, so it is used as a positive test indicator for many kinds of tumors. This paper reviews recent advances in the detection of the tumor marker AFP based on three immuno-biosensors: electrochemical (EC), photoelectrochemical (PEC), and electrochemical luminescence (ECL). The electrodes are modified by different materials or homemade composites, different signaling molecules are selected as single probes or dual probes for the detection of AFP. The detection limit was as low as 3 fg/mL, which indicated that the AFP immunosensor had achieved highly sensitive detection. In addition, we also reviewed and summarized the current development status and application prospect of AFP immunoelectrochemical sensors. There are not too many researches on immunosensors based on dual-signal ratios, and the commonly used probes are methylene blue (MB) and ferrocene (Fc). It would be more innovative to have more novel signaling molecules as probes to prepare dual-signal ratio sensors.

Palladium-Copper Bimetallic Aerogel as New Modifier for Highly Sensitive Determination of Bisphenol A in Real Samples.

In this study, a bimetallic palladium-copper aerogel was synthesized and used for modification of a graphite paste electrode (Pd-Cu/GPE), allowing the sensitive determination of bisphenol A (BPA). Different techniques, such as SEM, TEM, XPS, and AFM, were used for characterization of the Pd-Cu aerogel. To elucidate the properties of the Pd-Cu/GPE, the electrochemistry methods such as differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy were used. DPV measurements were conducted in phosphate electrolyte and buffer solution (0.2 M PBS, pH 5) at a potential range from 0.4 to 0.9 V vs. Ag/AgCl. The DPVs peaks currents increased linearly with BPA concentrations in the 0.04-85 and 85-305 µM ranges, with a limit of detection of 20 nM. The modified electrode was successfully used in real samples to determine BPA, and the results were compared to the standard HPLC method. The results showed that the Pd-Cu/GPE had good selectivity, stability, and sensitivity for BPA determination.

Electrochemical sensor for ultrasensitive sensing of biotin based on heme conjugated with gold nanoparticles and its electrooxidation mechanism.

We report the fabrication of a facile sensor using heme conjugated with gold nanoparticles (AuNPs) in situ on a glass carbon electrode (GCE) for the ultrasensitive determination of biotin without antibody or streptavidin. The use of heme and AuNPs as dual amplifiers allows a very broad detection range from 0.0050 to 50.0000 µmol·L-1 and a very low detection limit of 0.0016 µmol·L-1. The mechanistic aspects were elucidated using electrochemical analyses and frontier orbital calculations showing that the electrooxidation of biotin involves a one-electron and a one-proton transfer, generating biotin sulfoxide. The heme/AuNPs/GCE sensor exhibited excellent selectivity, reproducibility and stability, indicating high robustness. The recovery was between 97.20 and 105.70% with RSD less than 8.71%, suggesting good practicability. Our studies demonstrate that this approach can be used to detect and quantify biotin in a range of foods, including milk, infant formula, flour, orange juice, mango juice, egg white and egg yolk. Furthermore, all measurements do not require any intricate preparation or pre-treatment of the foods, thus representing a great potential for point-of-care testing.

A facile indole probe for ultrasensitive immunosensor fabrication toward C-reactive protein sensing.

C-reactive protein (CRP) is a protein biomarker for acute phase response. Herein, we fabricate a highly sensitive electrochemical immunosensor for CRP on a screen-printed carbon electrode (SPCE) with indole as a novel electrochemical probe and Au nanoparticles for signal amplification. Amongst, indole appeared as transparent nanofilms on the electrode surface, and underwent a one-electron and one-proton transfer to form oxindole during the oxidation process. Upon optimization of experimental conditions, a logarithmic correlation between CRP concentration (0.0001-100 µg∙mL-1) and response current was revealed with a detection limit of 0.03 ng∙mL-1 and a sensitivity of 5.7055 µA∙µg-1∙mL∙cm-2. The sensor exhibited exceptionally distinction selectivity, reproducibility and stability of the electrochemical immunosensor studied. The recovery rate of CRP in human serum samples determined by the standard addition method, ranged between 98.2-102.2%. Overall, the developed immunosensor is promising, and has the potential for CRP detection in real human serum samples.

Peptide Hydrogels as Immunomaterials and Their Use in Cancer Immunotherapy Delivery.

Peptide-based hydrogel biomaterials have emerged as an excellent strategy for immune system modulation. Peptide-based hydrogels are supramolecular materials that self-assemble into various nanostructures through various interactive forces (i.e., hydrogen bonding and hydrophobic interactions) and respond to microenvironmental stimuli (i.e., pH, temperature). While they have been reported in numerous biomedical applications, they have recently been deemed promising candidates to improve the efficacy of cancer immunotherapies and treatments. Immunotherapies seek to harness the body's immune system to preemptively protect against and treat various diseases, such as cancer. However, their low efficacy rates result in limited patient responses to treatment. Here, the immunomaterial's potential to improve these efficacy rates by either functioning as immune stimulators through direct immune system interactions and/or delivering a range of immune agents is highlighted. The chemical and physical properties of these peptide-based materials that lead to immuno modulation and how one may design a system to achieve desired immune responses in a controllable manner are discussed. Works in the literature that reports peptide hydrogels as adjuvant systems and for the delivery of immunotherapies are highlighted. Finally, the future trends and possible developments based on peptide hydrogels for cancer immunotherapy applications are discussed.

Microencapsulation of Probiotic Streptococcus salivarius LAB813.

Probiotics are living microorganisms that confer a health benefit on the host when administered in adequate amounts. Streptococcus salivarius, a commensal bacterium found in the oral cavity, has been shown to secrete antimicrobial peptides and can be used as probiotics. This study aimed to develop a delivery system for the probiotic LAB813, a novel S. salivarius strain first identified in the laboratory. Probiotics can be delivered and protected through the encapsulation of biomaterials such as polysaccharides. Their biocompatibility, biodegradability, user-friendliness, and ease of access make polysaccharides useful for encapsulating probiotics. Alginate (Alg) and chitosan (Ch) are naturally obtained polysaccharides and, hence, tested for LAB813 encapsulation. An extrusion method of encapsulation was performed to form Alg microcapsules (Alg-LAB813), some of which were coated with Ch (Alg-LAB813-Ch) to provide dual-layered protection. Inhibitory assays of the Alg-LAB813 and Alg-LAB813-Ch microcapsules were assayed against an indicator strain. Alg-LAB813-Ch microcapsules showed superior antibacterial properties compared to Alg-LAB813 microcapsules over 24 h and when subject to temperatures ranging from 4 to 68 °C. In addition, Alg-LAB813-Ch microcapsules retained antibacterial activity for up to 28 days of storage at 4 °C. The strong and sustained inhibitory activities of Ch-coated Alg encapsulated LAB813 signify the potential for their use to improve oral health.

Facile bimetallic co-amplified electrochemical sensor for folic acid sensing based on CoNPs and CuNPs.

Folic acid (FA) is essential for human health, particularly for pregnant women and infants. In this work, a glassy carbon electrode (GCE) was modified by a bimetallic layer of Cu/Co nanoparticles (CuNPs/CoNPs) as a synergistic amplification element by simple step-by-step electrodeposition, and was used for sensitive detection of FA. The proposed CuNPs/CoNPs/GCE sensor was characterized by differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS) and field emission scanning electron microscopy (FESEM). Then, under optimal conditions, a linear relationship was obtained in the wide range of 110.00-1750.00 µM for the detection of FA with a limit of detection (LOD) of 34.79 µM (S/N = 3). The sensitivity was calculated as 0.096 µA µM-1 cm-2. Some interfering compounds including glucose (Glc), biotin, dopamine (DA), and glutamic acid (Glu) showed little effect on the detection of FA by amperometry (i-t). Finally, the average recovery obtained was in a range of 91.77-110.06%, with a relative standard deviation (RSD) less than 8.00% in FA tablets, indicating that the proposed sensor can accurately and effectively detect the FA content in FA tablets.

Development of a Bacterial Enzyme-Based Biosensor for the Detection and Quantification of Selenate.

An enzymatic biosensor has been developed for the determination of selenate (SeO4 2- ), in which selenate reductase (SeR) is chemically attached to a gold disk electrode by lipoic acid N-hydroxysuccinimide ester as linker, allowing the catalytic reduction of the SeO4 2- to SeO3 2- . Modification of the gold electrode was characterized by X-ray photoelectron spectroscopy (XPS), time-of-flight secondary ion mass spectroscopy (ToF-SIMS), and electrochemistry. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) measurements were performed in different buffers for selenate determination. Under optimum conditions, the calibration curve was linear over the range 7.0-3900.0 µg L-1 with limits of detection and quantification of 4.97 and 15.56 µg L-1 , respectively. The possible interference of the relevant oxyanions SO4 2- , NO3 - , NO2 - , PO4 3- and AsO4 3- in the determination of SeO4 2- was studied. Finally, the proposed biosensor was used to determine SeO4 2- with recovery between 95.2 and 102.4 % in different real water samples.

Enzyme-free glucose sensors with efficient synergistic electro-catalysis based on a ferrocene derivative and two metal nanoparticles.

Gold electrodes (GE) were modified by the deposition of copper nanoparticles (CuNPs) and cobalt nanoparticles (CoNPs), followed by drop-casting of the ferrocene derivative FcCO-Glu-Cys-Gly-OH (Fc-ECG), resulting in two enzyme-free electrochemical sensors Fc-ECG/CuNPs/GE and Fc-ECG/CuNPs/GE. The ferrocene-peptide conjugate acts as an effective redox mediator for glucose oxidation, while metal nanoparticles acted as non-biological sites for glucose oxidation. Field emission scanning electron microscopy (FESEM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were carried out for characterization, while differential pulse voltammetry (DPV) was used for glucose quantification. Under optimized conditions, DPV shows a linear relationship between glucose concentration and the peak current. Both sensors showed a surprisingly high sensitivity of 217.27 and 378.70 µA mM-1 cm-2, respectively. A comparison to other glucose sensors shows a sensitivity that is 25 times higher. The sensors exhibit good reproducibility, stability, and repeatability. In injection experiments, recovery rates were 87.39-107.65% and 100.00-106.88%, respectively.

One step construction of an electrochemical sensor for melamine detection in milk towards an integrated portable system.

Excessive intake of melamine (MEL) can be harmful to human health, and it is important to establish a rapid and accurate MEL detection method. As the electrochemical activity of MEL is very low, ferrocenylglutathione (Fc-ECG) was used as an electron transfer mediator to assist with the detection of MEL using screen-printed carbon electrode (SPCE). This modified electrode (MEL/Fc-ECG/SPCE) was prepared by stepwise drop-casting and was fully characterized. Results showed that MEL significantly enhanced signal of Fc-ECG/SPCE sensor due to the three p-π conjugated double bonds that facilitated electron transfer. Under optimal conditions, the sensor exhibits two linearities in the range of 0.20-2.00 µM and 8.00-800 µM, with a sensitivity of 15.03 µA·µM-1·cm-2. The selectivity, stability, and reproducibility were investigated and successfully used to detect MEL in raw milk and confirms safety verification of foods. Moreover, a portable testing platform was designed for MEL detection based on a CH32 chip.

Metal-dependent electrochemical discrimination of DNA quadruplex sequences.

Films of four different DNA quadruplex-forming (G4) sequences (c-KIT, c-MYC, HTelo, and BCL2) on gold surfaces were investigated by electrochemical impedance spectroscopy (EIS) to evaluate whether they evoke unique electrochemical responses that can be used for their identification. This could render EIS an alternative means for the determination of G4 sequences of unknown structure. Towards, this end, cation-dependent topology changes in the presence of either K+, K+ in combination with Li+, or Pb2+ in the presence of Li+ were first evaluated by circular dichroism (CD) spectroscopy, and electrochemical studies were performed subsequently. As a result, G4-sequence specific charge transfer resistance (RCT) patterns were in fact observed for each G4 sequence, allowing their discrimination by EIS.

Nanoporous Gold for the Miniaturization of In Vivo Electrochemical Aptamer-Based Sensors.

Electrochemical aptamer-based sensors enable real-time molecular measurements in the living body. The spatial resolution of these measurements and ability to perform measurements in targeted locations, however, is limited by the length and width of the device's working electrode. Historically, achieving good signal to noise in the complex, noisy in vivo environment has required working electrode lengths of 3-6 mm. To enable sensor miniaturization, here we have enhanced the signaling current obtained for a sensor of given macroscopic dimensions by increasing its surface area. Specifically, we produced nanoporous gold via an electrochemical alloying/dealloying technique to increase the microscopic surface area of our working electrodes by up to 100-fold. Using this approach, we have miniaturized in vivo electrochemical aptamer-based (EAB) sensors (here using sensors against the antibiotic, vancomycin) by a factor of 6 while retaining sensor signal and response times. Conveniently, the fabrication of nanoporous gold is simple, parallelizable, and compatible with both two- and three-dimensional electrode architectures, suggesting that it may be of value to a range of electrochemical biosensor applications.

Development of a novel label-free impedimetric electrochemical sensor based on hydrogel/chitosan for the detection of ochratoxin A.

Ochratoxin A (OTA) is one of the most abundant mycotoxins that contaminate various food products. Herein, we propose a novel label-free impedimetric electrochemical sensor consisting of chitosan/dipeptide nanofibrous hydrogel and immobilized DNA probes with OTA aptamer for the detection of OTA. The thin film of chitosan/dipeptide nanofibrous hydrogel was used as sensing interface and carrier for hybridization chain reaction (HCR) of OTA aptamer and DNA2 strand to form DNA concatemer. The concatemer was dissociated to single-stranded DNA (ssDNA) in the presence of target OTA, and the signal amplification was further implemented by introducing RecJf exonuclease, which could digest the single-stranded DNA resulting in OTA recycle. Electrochemical impedance spectroscopy (EIS) has been employed to characterize the properties of the fabricated sensor. A linear detection range of 0.1-100 ng mL-1 was obtained for OTA with a low detection limit of 0.03 ng mL-1. Furthermore, the developed sensor was demonstrated in white wine to detect OTA, indicating that the proposed impedimetric sensor has a promising potential application in the food industry.

Consecutive Silver(I) Ion Incorporation into Oligonucleotides containing Cytosine-Cytosine Mispairs.

Invited for this month's cover is the group of Prof. Heinz-Bernhard Kraatz from the University of Toronto, Canada. The cover picture shows the consecutive incorporation of AgI ions into a DNA duplex containing adjacent C-C mispairs. Read the full text of the article at 10.1002/cplu.202000607.

Electrochemical Reduction of CO2 at Coinage Metal Nanodendrites in Aqueous Ethanolamine.

Electrocatalytic reduction of CO2 into usable chemicals is a promising path to address climate change and energy challenges. Herein, we demonstrate the synthesis of unique coinage metal (Cu, Ag, and Au) nanodendrites (NDs) via a facile galvanic replacement reaction (GRR), which can be effective electrocatalysts for the reduction of CO2 in an ethanolamine (EA) solution. Each metal ND surface was directly grown on glassy-carbon (GC) substrates from a mixture of Zn dust and the respective precursor solution. The electrocatalytic activities of the synthesized ND surfaces were optimized for CO2 reduction in EA solution by varying their composition. It was determined that a 0.05 mol fraction of EA exhibited the highest catalytic activity for all metal NDs. Linear sweep voltammetry (LSV) and electrochemical impedance spectroscopy (EIS) techniques showed that metal-ND electrodes possessed higher current densities, lower onset potentials and lower charge-transfer resistances for CO2 reduction than their smooth polycrystalline electrode counterparts, indicating improved CO2 reduction catalytic activity. It was determined, using FTIR and NMR spectroscopy, that formate was produced as a result of the CO2 reduction.

Peptide-Polydopamine Nanocomposite Hydrogel for a Laser-Controlled Hydrophobic Drug Delivery.

Smart antibacterial systems, delivering antimicrobials in a highly controlled manner, are one strategy toward fighting the rise of antibiotic-resistant pathogens. Here, we engineer a laser-responsive antimicrobial nanocomposite hydrogel combining a peptide amphiphile and a photothermally active polydopamine nanoparticle (PDNP) to entrap the hydrophobic rifampicin within the hydrophilic hydrogel matrix. We show that the ability of the gelator to interact and retain rifampicin within the gel induced structural changes in its nanofiber network and mechanical properties. Furthermore, PDNP inclusion enabled laser-induced drug release, preventing growth of a Gram-negative E. coli. Overall, our work provides a significant advance in designing smart materials for controlled drug delivery applications.

A novel electrochemical immunosensor for hepatitis B surface antigen based on Fe3O4 nanoflowers and heterogeneous chain reaction signal amplification strategy.

Herein, a novel sandwich-type electrochemical immunosensor was fabricated based on Fe3O4 nanoflowers (Fe3O4 NFs) and heterogeneous chain reaction (HCR) signal amplification strategy for the sensitive detection of hepatitis B surface antigen (HBsAg). The aldehyde-functionalized Fe3O4 NFs are used as a supporting matrix to immobilize the hepatitis B surface antibody 1 (HBsAb1). The biotin-modified single-strand DNA (biotin-S0) was connected onto the biotin-HBsAb2 via linkage of streptavidin (SA), followed by addition of methylene blue (MB) modified single strand DNA1 (MB-S1) and DNA2 (MB-S2) for HCR signal amplification. The designed immunosensor exhibited a detection linear range of 0.5 pg mL-1-0.25 ng mL-1 and a low detection limit of 0.16 pg mL-1, with excellent stability, selectivity and reproducibility. Furthermore, HBsAg is detected in the serum samples with a stable and fast response, indicating that the proposed immunosensor has a promising potential application in clinical analysis.