Perspectives and trends in advanced optical and electrochemical biosensors based on engineered peptides.

With the advancement of life medicine, in vitro diagnostics (IVD) technology has become an auxiliary tool for early diagnosis of diseases. However, biosensors for IVD now face some disadvantages such as poor targeting, significant antifouling properties, low density of recognized molecules, and poor stability. In recent years, peptides have been demonstrated to have various functions in unnatural biological systems, such as targeting properties, antifouling properties, and self-assembly properties, which indicates that peptides can be engineered. These properties of peptides, combined with their good biocompatibility, can be well applied to the design of biosensors to solve the problems mentioned above. This review provides an overview of the properties of engineered functional peptides and their applications in enhancing biosensor performance, mainly in the field of optics and electrochemistry.

Ti3C2Tx MXene -facilitated non-selective trapping effect: Efficient SERS detection of exosomal PD-L1.

Biosensors developed through a sandwich approach have demonstrated favorable detection performance for exosomal programmed cell death 1 ligand 1 (ExoPD-L1) detection. However, the reported PD-L1 antibodies, peptides, and aptamers utilized in these biosensors typically bind to the extracellular region, with overlapping binding sites that severely constrain the fabrication of biosensors. In this study, we present a simple approach to specifically identify and analyze ExoPD-L1 through the non-selective trapping effect of Ti3C2TX (X=-O, -F, -OH) MXene on exosomes via the formation of Ti-O-P complexation, and the selective capture of peptide-functionalized Au@MPBA (4-Mercaptophenylboronic acid) @SiO2 surface enhanced Raman scattering (SERS) tags on ExoPD-L1. The biosensor delivered a both hypersensitive and reliable performance in exosome detection with a low limit of detection (20.74 particles/mL) in the linear range of 102 to 5×106 particles/mL. Furthermore, the biosensor demonstrated excellent stability and interference resistance in detecting ExoPD-L1 in clinical serum samples, enabling the easy differentiation of breast cancer patients from healthy controls. This work provides new insights into the design of biosensors for exosome detection and can serve as a replicable template for sandwich immunoassay detection for other types of sensors, including but not limited to SERS.

Tunable Au@SiO2/Au Film Metasurface as Surface Plasmon Resonance Enhancer for Direct and Ultrasensitive Detection of Exosomes.

Surface plasmon resonance (SPR) spectroscopy with non-labelling, sensitive, and real-time properties is critical for clinical diagnosis applications. However, conventional SPR sensors face the challenge of lower sensitivity and selectivity for trace exosomes assay in complex serum. We proposed a core-shell Au@SiO2-Au film (Au@SiO2-Au film) metasurface to enhance SPR signal based on systematic study on the relationship between gap modes and SPR enhancement. The self-assembled multifunctional peptide was designed as recognition layer with antifouling properties for ultrasensitive and selective detection of PD-L1+ exosomes in serum. The tuning electromagnetic (EM) field model by manipulating the gap was established to guide the preparation of Au@SiO2-Au film metasurface. The in-plane and out-of-plane coupling of Au@SiO2 nanoparticles (NPs) could greatly enlarge and enhance three-dimensional EM field to meet the size of exosomes located in the evanescent field. At the structural level, we achieved high sensitivity (0.16 particles/mL) and a broad response range (10-5 × 103 particles/mL) through optimizing the thickness of SiO2 and surface coverage of Au@SiO2. Furthermore, clinical sample assay achieved the optimal diagnostic accuracy (AUC = 0.97) for differentiating cancer patients from healthy controls. This work provides an opportunity for the construction of a tunable gap mode as SPR enhancer in a total internal reflection architecture. The systematic study on the relationship between gap modes and SPR sensitivity provides a broad scope for promoting direct, efficient, highly selective, and sensitive detection of SPR sensors for clinical application.

2D MOF-enhanced SPR sensing platform: Facile and ultrasensitive detection of Sulfamethazine via supramolecular probe.

Sulfamethazine (SMZ) is widely present in the environment and can cause severe allergic reactions and cancer in humans. Accurate and facile monitoring of SMZ is crucial for maintaining environmental safety, ecological balance, and human health. In this work, a real-time and label-free surface plasmon resonance (SPR) sensor was devised using a two-dimensional metal-organic framework with superior photoelectric performance as an SPR sensitizer. The supramolecular probe was incorporated at the sensing interface, allowing for the specific capture of SMZ from other analogous antibiotics through host-guest recognition. The intrinsic mechanism of the specific interaction of the supramolecular probe-SMZ was elucidated through the SPR selectivity test in combination with analysis by density functional theory, including p-π conjugation, size effect, electrostatic interaction, π-π stacking, and hydrophobic interaction. This method facilitates a facile and ultrasensitive detection of SMZ with a limit of detection of 75.54 pM. The accurate detection of SMZ in six environmental samples demonstrates the potential practical application of the sensor. Leveraging the specific recognition of supramolecular probes, this direct and simple approach offers a novel pathway for the development of novel SPR biosensors with outstanding sensitivity.

Current methods and emerging approaches for detection of programmed death ligand 1.

Various tumor cells overexpress programmed death ligand 1 (PD-L1), a main immune checkpoint protein (ICP) embedded in the tumor cells membrane, to evade immune recognition through the interaction between PD-L1 and its receptor programmed death 1 (PD-1) which is from T-cells for maintaining immune tolerance. So inhibitors targeting the PD-1 or PD-L1 can block the PD-1/PD-L1 signaling pathway to restore the recognition activity of the immune system to tumor cells, which also have been utilized as a novel approach to improve the clinical therapeutic effect for cancer patients. Since not all cancer patients can respond to these inhibitors effectively, previous diagnosis of PD-L1 is significant to target the right treatments for cancer patients. This review pays attention to the PD-L1 detection and recent progress in the measurement of PD-L1 concentration, including various detection methods based on optical sensors as well as electrochemical assays. Apart from above those, we also focus on the prospects of PD-L1 detection in precision medicine.

Antibacterial Activity of Graphene-Based Nanomaterials.

Recent research has shown that graphene as a novel "green" antibacterial material possess excellent antibacterial properties with no risk of bacterial resistance for daily life due to its physical damage-based bactericidal mechanism. Therefore, an increasing amount of research has been focused towards evaluating the antibacterial effects of graphene and graphene-based hybrid materials. In this chapter, we reviewed the antibacterial activity and mechanism of graphene-based nanomaterials and highlighted the importance of size, morphology, and composites in the application of antibacterial materials development. Finally, we made a summary and outlook on this research field.

Aptamer-Assisted Protein Orientation on Silver Magnetic Nanoparticles: Application to Sensitive Leukocyte Cell-Derived Chemotaxin 2 Surface Plasmon Resonance Sensors.

Leukocyte cell-derived chemotaxin 2 (LECT2) has been proved to be a potential biomarker for the diagnosis of liver fibrosis. In this work, a sensitive surface plasmon resonance (SPR) assay for LECT2 analysis was developed. Tyrosine kinase with immune globulin-like and epidermal growth factor-like domains 1 (Tie1) is an orphan receptor of LECT2 with a C-terminal Fc tag, which is far away from the LECT2 binding sites. The Fc aptamer was intentionally used to capture the Tie1 through its Fc tag, connecting with Fe3O4-coated silver magnetic nanoparticles (Ag@MNPs) and ensuring the LECT2 binding site to be outward. Attributed to the orientation nature of the captured protein, Ag@MNPs were able to enhance the SPR signal. A sensitive LECT2 sensor was successfully fabricated with a detection limit of 10.93 pg/mL. The results showed that the immobilization method improved the binding efficiency of Tie1 protein. This strategy could be extended to attach antibodies or recombinant Fc label proteins to Fc aptamer-based nanoparticles.

Rapid and sensitive detection of PD-L1 exosomes using Cu-TCPP 2D MOF as a SPR sensitizer.

Two-dimensional metal organic framework (2D MOF Cu-TCPP) with significantly enhanced photoelectric properties was synthesized by a simple hydrothermal method. The π-stacked electroactive porphyrin molecules of TCPP-based 2D MOF carry out charge transport in the MOF structure. The d-d band transition of Cu2+ and its 2D ultra-thin characteristics can produce excellent near-infrared light absorption to couple with SPR. Three key parameters including the refractive index sensitivity, detection accuracy and quality factor were improved significantly for 2D MOF modified gold chips. Especially, the refractive index sensitivity was increased from 98 to 137.67°/RIU after modified with 2D MOF. Thus, for the first time, we applied it as a signal enhancer to improve direct SPR assay for the Programmed death ligand-1 (PD-L1) exosomes. Owning to its large specific surface area, excellent photoelectric properties, highly ordered structure, good dispersion and biocompatibility, the LOD of the SPR sensor was 16.7 particles/mL. The reliability and practicability were further validated by analysis of PD-L1 exosomes in human serum samples. The recovery rate was 93.43 %-102.35%, with RSD of 5.79 %-14.6%. Given their excellent signal amplification ability, 2D MOF Cu-TCPP could serve as an ideal SPR sensitizer for rapid and sensitive detection of trace disease markers.

The magnetic-nanoparticle-assisted sensitive detection of nitrated α-syn in blood based on a sensitizing electrochemical layer.

Parkinson's disease (PD) is the second most prevalent neurodegenerative disease. Nitrated α-synuclein (α-syn) in the blood is a potentially efficient biomarker for PD in its early stages. In this work, an ultrasensitive electrochemical immunosensor was developed for the specific detection of nitrated α-syn. Supramolecule-mediated AuNP composites (GNCs) were modified on the gold electrode as a sensing film to capture anti-nitrated α-syn. Basic characterization studies revealed that GNCs were composed of abundant binding sites and had high conductivity with a large surface area, biocompatibility, and remarkable electrochemical activity. Anti-α-syn-modified magnetic nanoparticles (MNPs) were used as signal amplification tags to construct a sensitive sandwich assay. With a high specific surface area, strong conductivity, and abundant active sites, GNCs as an amplifying matrix can enhance the performance of the immunoassay and obtain preliminary signal amplification. MNPs showed excellent stability and led to a net decrease in the charge-transfer resistance due to their unique spherical structure and high conductivity, resulting in a sensitive electrochemical signal change according to the nitrated α-syn concentration in the sample. Therefore, this simple nitrated α-syn immunoassay with sensitivity and selectivity has potential for practical clinical applications.

Pyridinium porphyrins and AuNPs mediated bionetworks as SPR signal amplification tags for the ultrasensitive assay of brain natriuretic peptide.

Extension of the self-assembled bionanonetworks into surface plasmon resonance (SPR) assay investigation provides an effective signal amplification approach. We fabricated a bionetwork by nucleic acids, organic compounds, and supramolecular gold nanoparticles for ultrasensitive SPR detection of B-type natriuretic peptide (BNP). The SPR method was developed by a sandwich-type format of aptamer-target-antibody, and the aptamer-modified bionanonetworks induced localized SPR and large refractive index for different concentrations of the target BNP. The linear concentration range and limit of detection were 1-10,000 pg/mL (R2 = 0.9852) and 0.3 pg/mL respectively. The detection recovery was in the range 92.13 to 108.69%. The approach embraces the following main advantages: (1) Cooperative double recognition was realized by calix[4]arenes for amino aptamers and pyridinium porphyrins. (2) The approach provided the specificity for supramolecular-based nanomaterials and a simple synthesis process via the ordered self-assembly under mild conditions. (3) The bionanonetworks endowed the SPR assay with signal amplification and stable determination for trace proteins. Therefore, it is expected that this study may offer a new SPR signal-amplified platform of organic-inorganic bionanonetworks to achieve sensitive, stable, and real-time determination. Graphical abstract Schematic of bionanonetwork based on porphyrin-mediated functionalized gold nanoparticles for SPR signal amplification to quantitatively detect BNP.

Inorganic and Metal-Organic Nanocomposites for Cascade-Responsive Imaging and Photochemical Synergistic Effects.

Porphyrins coordinated with platinum(II) chemotherapeutic drugs are attractive for the development of photosensitizers for photodynamic therapy (PDT) of cancer. In this paper, inorganic and metal-organic nanocomposites were synthesized with cascade-responsive imaging and photochemical synergistic effects. After endo/lysosomal escape, the outer metal-organic frameworks were degraded, leading to the release of an excellent photosensitizer (tetrapyridylporphyrin, PtTPyP). Subsequently, doxorubicin (DOX), inserted in the adenosine triphosphate (ATP) aptamer-functionalized gold nanoparticles, was released under the stimulation of endogenous ATP, synergistically enhancing cancer treatment. Fluorescence imaging allowed tracking of PtTPyP and DOX for real-time detection and on-demand therapy. This strategy endowed the nanocomposites with stability, responsiveness, effectiveness, and ease of synthesis, namely, sTREE strategy. Accordingly, our demonstration provided a promising and smart nanocarrier for imaging and drug delivery.

A facile gold nanoparticles embeded hydrogel for non-enzymatic sensing of glucose.

The assembly of nanoparticle into electrodes with precise structure and uniform core sizes is important for electrocatalysis. In this study, we reported on a simple strategy for in-situ preparation of gold nanoparticles embedded D-sorbitol hydrogel (D-gel@AuNPs). D-sorbitol hydrogel with acyl hydrazide (D-gel) was synthesized and characterized. AuNP's stable electronic structure, high surface coverage and good conductivity was achieved enabled D-gel@AuNPs exhibits the enhanced electrocatalytic performance. The electrochemical results reveal that the catalytic progress is highly promoted by the D-gel@AuNPs with a detection limit of 0.067 mM and detection range of 0.1-30 mM. The high enzymatic activity and stability provide the high possibility for the development of high value glucose sensors. This mechanistically novel strategy expands the scope of assembly of NPs method for the development of enhanced other electrochemical properties such as amperometric sensing and photcatalysis applications, as well as electrocatalysis.

Analyte-resolved magnetoplasmonic nanocomposite to enhance SPR signals and dual recognition strategy for detection of BNP in serum samples.

B-type natriuretic peptide (BNP) is a short peptide that is considered to be an important heart failure (HF)-related biomarker. Due to its low concentration in the blood and short half-life, the sensitive detection of BNP is a bottleneck for diagnosing patients at early stages of HF. In this paper, we report a facile surface plasmon resonance (SPR) sensor to measure BNP; the sensor is based on aptamer-functionalized Au nanoparticles (GNPs-Apt) and antibody-modified magnetoplasmonic nanoparticles (MNPs-Ab) to enable dual screening of BNP in complex environments. During sensing, BNP forms MNP-Ab/BNP/GNP-Apt nanoconjugates that can be rapidly separated from the complex sample by a magnet to avoid degradation within the analyte's half-life. The developed SPR biosensor shows high selectivity, a wide dynamic response range of BNP concentrations from 100 fg/mL to 10 ng/mL, and a low detection limit of 28.2 fg/mL (S/N = 3). Using the proposed sensor, BNP was successfully detected in clinical samples. Thus, the designed SPR biosensor provides a novel and sensitive sensing platform for BNP detection with potential applications in clinical practice.

Analyte induced AuNPs aggregation enhanced surface plasmon resonance for sensitive detection of paraquat.

Paraquat (PQ) residue is harmful for people's health. This work fabricated an efficient approach to determine PQ sensitively. We exploited a novel surface plasmon resonance (SPR) detection system based on the analyte induced network architecture of supermolecules modified gold nanoparticles (AuNPs) on the chip surface. para-Sulfonatocalix[4]arene (pSC4) were used as a recognition molecule for paraquat. PQ can mediate the aggregation of pSC4 capped AuNPs (pSC4-AuNPs) through the host-guest recognition, which can be used as signal amplification for PQ assay. This achievement is due to several outstanding properties of this detection system: first, local SPR and high refractive index of AuNPs can enhance the signal of SPR dramatically; second, AuNPs is more stable and biocompatible and diffusely used in colorimetric methods; third, the network AuNPs structure has unique photo characterization for enhancement of SPR. Analyte induced AuNPs aggregation amplified SPR assay shows dramatic signal enhancement ability. The detection limit for PQ was found to be 2.2 pM This strategy provides a new concept for developing sensitive SPR sensors for the highly selective detection of small molecules.

para-Sulfonatocalix[4]arene stabilized gold nanoparticles multilayers interfaced to electrodes through host-guest interaction for sensitive ErbB2 detection.

Nanoparticle (NP) structure, compositing and the nature of the NP-functionalized electrode interface have a strong influence upon electrochemical properties that are critical in applications such as sensing, photocatalysis and electrocatalysis. Existing methods to fabricate NP-functionalized electrodes do not allow or precise control over all these variables, especially the NP-electrode interface, making it difficult to understand and predict how structural changes influence electrode activity and consequently limit the application. To conquer this problem, in this study, we fabricated a stepwise construction of a novel supramolecular stabilized gold nanoparticles (AuNPs) multilayer mediated by guest molecules, yielding 3D AuNPs assembly at the electrode interface. para-Sulfonatocalix[4]arene (pSC4), a water soluble macrocyclic synthetic receptor, has been served as a stabilizing ligand for preparation and gaining new insights into pSC4 stabilized gold nanoparticles (pSC4-AuNPs) tethered on the electrode interface through host-guest interaction. We investigated the electrochemical properties of multilayer pSC4-AuNPs modified gold electrode using different core size of AuNPs with varying layer number. The electron transfer ability was characterized by electrochemical impedance spectroscopy (EIS). Electrochemical signals are significantly enhanced through the layer-by-layer assembly of pSC4-AuNPs due to its high conductivity and high effective area. With this innovative method, by taking the assay of a tumor marker as an example, human epidermal growth factor receptor 2 (ErbB2) was successfully measured with a detection limit of 0.5ng/mL. Taking the advantage of the pSC4-AuNPs multilayer's good biocompatibility, high effective area and high electronic transmission, 3D AuNPs multilayer produced on the electrode interface suggests a portable synthetic pathway for the application into sensitive electrochemical biosensor.

CB[7]-mediated signal amplification approach for sensitive surface plasmon resonance spectroscopy.

Cucurbit[7]uril (CB[7]) has received increasing attention because of its unique structure and multiple recognition properties. To improve the sensitivity of surface plasmon resonance (SPR) biosensors, we designed a novel strategy in which caspase-3 serves as the model analyte and CB[7]-modified AuNPs (CB[7]-AuNPs) act as the intermedium. The substrate peptides can be cleaved and replaced with a new N-terminal Phe residue in presence of caspase-3. The CB[7]-AuNPs combine with the N-terminal Phe on the gold chip surface through incorporating the side chain within the nonpolar CB[7] cavity and chelating the N-terminal ammonium group with CB[7] carbonyl oxygen. Then CB[7]-AuNPs integrate with short peptide-modified AuNPs containing Phe at the N-terminal of the peptide. SPR signals are significantly improved through the layer-by-layer assembly of AuNPs. The well-designed sensing platform allows the detection of caspase-3 in a linear range from 10fg/mL to 10(3)fg/mL with a detection limit of 2.2 fg/mL. Given its high specificity and desirable sensitivity, this CB[7]-assisted SPR method may be a useful tool for the assay of caspase-3 in the future. This work may also afford a new model to improve the sensitivity and selectivity of SPR biosensors in other protein detection experiments and disease diagnosis.

Evaluation of ß-Amyloid Peptides Fibrillation Induced by Nanomaterials Based on Molecular Dynamics and Surface Plasmon Resonance.

This report investigated the effect of carbon nanomaterials, single-wall carbon nanotube (SWCNT) and graphene oxide, on fibrillation of ß-amyloid 40 (Aß40) based on surface plasmon resonance (SPR) and molecular dynamics (MD). MD simulations are carried out in order to reveal the molecular mechanisms of the interaction between nanomaterials and Aß40. The strong interaction between Aß40 and nanomaterials is related to Van der Waals forces and the Coulomb force, inducing delicate manipulation of the main bonding energy for fibrillation of Aß40. The interaction energy between the Aß peptide and graphene is higher than that of SWCNT. Experimental results show both carbon nanomaterials enhance the appearance of a critical nucleus for nucleation of peptide fibrils. Graphene is more beneficial to assist the nucleation process than SWCNT. Combination of SPR and molecular dynamics could be a high-throughput method to screen protein fibrillation.

Dual-Mode SERS-Fluorescence Immunoassay Using Graphene Quantum Dot Labeling on One-Dimensional Aligned Magnetoplasmonic Nanoparticles.

A novel dual-mode immunoassay based on surface-enhanced Raman scattering (SERS) and fluorescence was designed using graphene quantum dot (GQD) labels to detect a tuberculosis (TB) antigen, CFP-10, via a newly developed sensing platform of linearly aligned magnetoplasmonic (MagPlas) nanoparticles (NPs). The GQDs were excellent bilabeling materials for simultaneous Raman scattering and photoluminescence (PL). The one-dimensional (1D) alignment of MagPlas NPs simplified the immunoassay process and enabled fast, enhanced signal transduction. With a sandwich-type immunoassay using dual-mode nanoprobes, both SERS signals and fluorescence images were recognized in a highly sensitive and selective manner with a detection limit of 0.0511 pg mL(-1).

Sensitive colorimetric assays for α-glucosidase activity and inhibitor screening based on unmodified gold nanoparticles.

A colorimetric sensor has been developed in this work to sensitively detect α-glucosidase activity and screen α-glucosidase inhibitors (AGIs) utilizing unmodified gold nanoparticles (AuNPs). The sensing strategy is based on triple-catalytic reaction triggered by α-glucosidase. In the presence of α-glucosidase, aggregation of AuNPs is prohibited due to the oxidation of cysteine to cystine in the system. However, with addition of AGIs, cysteine induced aggregation of AuNPs occurs. Thus, a new method for α-glucosidase activity detection and AGIs screening is developed by measuring the UV-vis absorption or visually distinguishing. A well linear relation is presented in a range of 0.0025-0.05 U mL(-1). The detection limit is found to be 0.001 U mL(-1) for α-glucosidase assay, which is one order of magnitude lower than other reports. The IC50 values of four kinds of inhibitors observed with this method are in accordance with other reports. The using of unmodified AuNPs in this work avoids the complicated and time-consuming modification procedure. This simple and efficient colorimetric method can also be extended to other enzymes assays.

Cultures of Schwann-like cells differentiated from adipose-derived stem cells on PDMS/MWNT sheets as a scaffold for peripheral nerve regeneration.

Carbon nanotubes (CNTs) are promising candidates as novel scaffolds for peripheral nerve regeneration. Schwann cells (SCs) are attractive therapeutic targets due to their pivotal role in peripheral nerve regeneration, but primary SCs have limitations for clinical application. However, adipose-derived stem cells (ASCs) may differentiate into Schwann-like cells. The present study assesses the potential applicability of multiwall CNTs (MWNTs) composited with polydimethylsiloxane (PDMS), which were then seeded with differentiated adipose-derived stem cells (dASCs) to promote neuronal differentiation and growth. Aqueous MWNT dispersion was filtered, and the PDMS/MWNT sheets were prepared using a simple printing-transfer method. Characterization of PDMS/MWNT sheets indicated their unique physical properties, such as superior mechanical strength and electroconductivity, compared with bare PDMS sheets. ASCs were differentiated into Schwann-like cells using a mixture of glial growth factors. Dorsal root ganglion (DRG) neurons were co-cultured with SCs and dASCs on PDMS/MWNTs sheets or noncoated dishes. An alamar blue proliferation assay of dASC and SCs showed significantly more dASC and SCs cultured on PDMS/MWNT sheets at 48 h and 72 h than when cultured on noncoated dishes (p < 0.05). Additionally, when DRG were cultured on PDMS/MWNT sheets seeded with dASCs, the proliferation of DRG neurons and the longest neurite outgrowth length per neuron were significantly greater than when DRG were cultured on PDMS/MWNT sheets alone or on noncoated dishes seeded with SCs or dASCs (p < 0.05). Overall, PDMS/MWNT sheets exhibited excellent biocompatibility for culturing Schwann-like cells differentiated from ASCs. Seeding the dASCs on PDMS/MWNT sheets may produce synergistic effects in peripheral nerve regeneration, similarly to SCs.