Specific Recognition Capability Research Articles

Magnetically Controlled Nanorobots Based on Red Emissive Peptide Dots and Artificial Antibodies for Specific Recognition and Smart Scavenging of Nε-(Carboxymethyl)lysine in Dairy Products

Food-borne advanced glycation end products (AGEs) are highly related to various irreversible diseases, and Nε-(carboxymethyl)lysine (CML) is the typical hazardous AGE. The development of feasible strategies to monitor and reduce CML exposure has become desirable to address the problems. In this work, we proposed magnetically controlled nanorobots by integrating an optosensing platform with specific recognition and binding capability, realizing specific anchoring and accurate determination as well as efficient scavenging of CML in dairy products. The artificial antibodies offered CML imprinted cavities for highly selective absorption, and the optosensing strategy was designed based on electron transfer from red emissive self-assembling peptide dots (r-SAPDs) to CML, which was responsible for the identity, response, and loading process. The r-SAPDs overcame the interference from autofluorescence, and the limit of detection was 0.29 μg L-1, which bestowed accuracy and reliability for in situ monitoring. The selective binding process was accomplished within 20 min with an adsorption capacity of 23.2 mg g-1. Through an external magnetic field, CML-loaded nanorobots were oriented, moved, and separated from the matrix, which enabled their scavenging effects and reusability. The fast stimuli-responsive performance and recyclability of the nanorobots provided a versatility strategy for effective detection and control of hazards in food.

Molecularly Imprinted Polymer-Based Sensors for Protein Detection.

The accurate detection of biological substances such as proteins has always been a hot topic in scientific research. Biomimetic sensors seek to imitate sensitive and selective mechanisms of biological systems and integrate these traits into applicable sensing platforms. Molecular imprinting technology has been extensively practiced in many domains, where it can produce various molecular recognition materials with specific recognition capabilities. Molecularly imprinted polymers (MIPs), dubbed plastic antibodies, are artificial receptors with high-affinity binding sites for a particular molecule or compound. MIPs for protein recognition are expected to have high affinity via numerous interactions between polymer matrices and multiple functional groups of the target protein. This critical review briefly describes recent advances in the synthesis, characterization, and application of MIP-based sensor platforms used to detect proteins.

Discrimination of cis-diol-containing molecules using fluorescent boronate affinity probes by principal component analysis.

Fluorescent boronate affinity molecules have gained increasing attention in the field of fluorescence sensing and detection due to their selective recognition capability towards cis-diol-containing molecules (cis-diols). However, the conventional fluorescent boronate affinity molecules face a challenge in differentiating the type of cis-diol only by their fluorescence responses. In this study, a simple method was used to discriminate different types of cis-diols, including nucleosides, nucleotides, sugars, and glycoproteins based on the phenylboronic acid-functionalized fluorescent molecules combined with principal component analysis (PCA). Both fluorescent molecules were simply synthesized by the covalent interaction between the amino group in 3-aminophenyl boronic acid and the isothiocyanate group in fluorescein or rhodamine B. In view of their fluorescence-responsive behaviors to these cis-diols directly, it is impossible to differentiate their types even under the optimized experimental conditions. When PCA was employed to treat the fluorescence response data and the quenching constants with their molecular weight, different types of cis-diols can be distinguished successfully. As a result, by integrating the fluorescence response of the boronate affinity probes with PCA, it can greatly improve the specific recognition capability of the boronic acids, providing a simple and direct way to distinguish and identify different types of cis-diols.

Early warning of Diaporthe infection in kiwifruit soft rot by plasmonic dimer-enhanced Raman spectroscopy.

Diaporthe genus is the dominant pathogens of kiwifruit soft rot with long incubation period and rapid onset and very hard to detect in advance. It is of great significance to develop point-of-care tests for disease prevention and field management. Here we screen a pair of genus-level primers for constructing plasmonic dimer Rayleigh/Raman spectroscopy, which enables rapid, specific, and sensitive detection of Diaporthe genus in real kiwifruits. Dark-filed Rayleigh scattering clearly visualizes the target-induced dimer assembly of nanoprobes. Plasmonic dimer-enhanced Raman spectroscopy has high specificity for 12 common isolates of Diaporthe genus in one batch samples with highly accurate results and much higher sensitivity than polymerase chain reaction (PCR). It realizes the recognition of Diaporthe infection at an early stage of 24h when the kiwifruits do not have any noticeable symptoms. It demonstrates a bright prospect of point-of-care tests for early warning of kiwifruit soft rot infection and quality control.

Facile and Compact Electrochemical Paper-Based Analytical Device for Point-of-Care Diagnostic of Dual Carcinogen Oxidative Stress Biomarkers through a Molecularly Imprinted Polymer Coated on Graphene Quantum-Dot Capped Gold.

Nanoscale imprinting significantly increases the specific surface area and recognition capabilities of a molecularly imprinted polymer by improving accessibility to analytes, binding kinetics, and template removal. Herein, we present a novel synthetic route for a dual molecularly imprinted polymer (dual-MIP) of the carcinogen oxidative stress biomarkers 3-nitrotyrosine (3-NT) and 4-nitroquinolin-N-oxide (4-NQO) as coatings on graphene quantum-dot capped gold nanoparticles (GQDs-AuNPs). The dual-MIP was successfully coated on the GQDs-AuNPs core via a (3-mercaptopropyl) trimethoxysilane (MPTMS) linkage and copolymerization with the 3-aminopropyltriethoxysilane (APTMS) functional monomer. In addition, we fabricated a facile and compact three-dimensional electrochemical paper-based analytical device (3D-ePAD) for the simultaneous determination of the dual biomarkers using a GQDs-AuNPs@dual-MIP-modified graphene electrode (GQDs-AuNPs@dual-MIP/SPGE). The developed dual-MIP device provides greatly enhanced electrochemical signal amplification due to the improved electrode-specific surface area, electrocatalytic activity, and the inclusion of large numbers of dual-imprinted sites for 3-NT and 4-NQO detection. Quantitative analysis used square wave voltammetry, with an oxidation current appearing at -0.10 V for 4-NQO and +0.78 V for 3-NT. The dual-MIP sensor revealed excellent linear dynamic ranges of 0.01 to 500 μM for 3-NT and 0.005 to 250 μM for 4-NQO, with detection limits in nanomolar levels for both biomarkers. Furthermore, the dual-MIP sensor for the simultaneous determination of 3-NT and 4-NQO provides high accuracy and precision, with no evidence of interference from urine, serum, or whole blood samples.

Novel Colorimetric Aptasensor for the Detection of Golgi Protein 73 by Exploiting the Peroxidase-Like Activity of the H-rGO-Mn3O4 Nanozyme

Golgi protein 73 (GP73) is a novel biomarker for the detection of hepatocellular carcinoma. We prepare a colorimetric aptasensor for GP73 detection based on the catalytic performance of the H-rGO-Mn3O4 nanozyme. The H-rGO-Mn3O4 nanozyme serves as a peroxidase mimetic which can enhance the reaction with the substrate (3, 3,’ 5, 5′-tetramethylbenzidine, TMB) and H2O2. The peroxidase-like activity of H-rGO-Mn3O4 features a 1.78-fold higher binding affinity value (Km) for TMB compared with that of horseradish peroxidase. The H-rGO-Mn3O4 nanozyme not only exhibits admirable peroxidase-like activities due to the synergistic effect of Mn3O4 NPs, rGO and hemin, but also has a large specific surface area to endow the GP73 aptamer with specific recognition capabilities. A sandwich colorimetric aptasensor is formed to realize the visual detection of GP73 through catalyzing the H2O2-mediated oxidation peroxidase substrate TMB to oxidized TMB accompanied by a color change from colorless to blue. Under optimal conditions, the logarithm of the GP73 concentration (0.05–50.0 ng/mL) shows a good linear relationship with the absorbance. The calibration equation is Y =−0.0383 lgC+0.4835, with R2 of 0.9964, and the lowest limit of detection is 36.94 pg/mL. Additionally, the content of GP73 in human serum samples is directly detected, and the relative standard deviation is 0.49–4.91%. Compared with the enzyme-linked immunosorbent assay method, the relative error is 0.23–3.61%. All in all, colorimetric aptasensor is demonstrated to exhibit excellent specificity, stability, and reproducibility.

Computer-aided design-based green fabrication of magnetic molecularly imprinted nanoparticles for specific extraction of non-steroidal anti-inflammatory drugs

Novel magnetic molecularly imprinted nanoparticles (MMINs) were greenly synthesized by one-pot method with ketoprofen (KP) as template molecule and used as the adsorbent of magnetic solid-phase extraction (MSPE) for selective extraction of non-steroidal anti-inflammatory drugs (NSAIDs). By means of calculation simulation technology, N-vinyl pyrrolidone (NVP) and allylglycine (AG) were rapidly screened as suitable dual functional monomers. After that, self-assembly solution of KP-NVP/AG, magnetic source (Fe3+/Fe2+) and N,N-dimethylformamide were mixed in one pot, and then simple hydrothermal technology was utilized to quickly prepare MMINs with a total reaction period of 3.5 h. A series of characterized techniques were adopted to study the structure, morphology and magnetic property of prepared MMINs. Results indicated that the adsorption behavior of MMINs towards KP was well fit for Freundlich adsorption model and pseudo-second-order kinetics. Under the optimal conditions, the obtained MMINs exhibited satisfactory specific recognition capability. The imprinting factor and extraction capacity towards KP were as high as 6.73 and 15.6 mg/g, respectively. Combining with HPLC, accurate and reliable approach for the measurement of trace NSAIDs in environmental waters and human urine samples was developed. The established approach exhibited low limits of detection (0.0024–0.0029 and 0.0026–0.0045 μg/L for water and urine samples, respectively), good fortified recoveries (77.8–119 % for water samples and 78.3–120 % for urine samples) and satisfactory precision (RSDs ≤ 9.7 %).

Optimization of Nanosubstrates toward Molecularly Surface-Functionalized Raman Spectroscopy.

Diagnostic advancements require continuous developments of reliable analytical sensors, which can simultaneously fulfill many criteria, including high sensitivity and specificity for a broad range of target analytes. Incorporating the highly sensitive attributes of surface-enhanced Raman spectroscopy (SERS) combined with highly specific analyte recognition capabilities via molecular surface functionalization could address major challenges in molecular diagnostics and analytical spectroscopy fields. Herein, we have established a controllable molecular surface functionalization process for a series of textured gold surfaces. To create the molecularly surface-functionalized SERS platforms, self-assembled benzyl-terminated and benzoboroxole-terminated monolayers were used to compare which thicknesses and root-mean-square (RMS) roughness of planar gold produced the most sensitive and specific surfaces. Optimal functionalization was identified at 80 ± 8 nm thickness and 7.2 ± 1.0 nm RMS. These exhibited a considerably higher SERS signal (70-fold) and improved sensitivity for polysaccharides when analyzed using principal component analysis (PCA) and self-organizing maps (SOM). These findings lay the procedure for establishing the optimal substrate specifications as an essential prerequisite for future studies aiming at developing the feasibility of molecular imprinting for SERS diagnostic applications and the subsequent delivery of advanced, highly selective, and sensitive sensing devices and analytical platforms.

Single-chain fragment variable produced by phage display technology: Construction, selection, mutation, expression, and recent applications in food safety.

Immunoassays are reliable, efficient, and accurate methods for the analysis of small-molecule harmful substances (such as pesticides, veterinary drugs, and biological toxins) that may be present in food. However, traditional polyclonal and monoclonal antibodies are limited by animal hosts and hinder further development of immunoassays. With the gradual application of phage display technology as an efficient in vitro selection technology, the single-chain fragment variable (scFv) now provides an exciting alternative to traditional antibodies. Efficiently constructed scFv source libraries and specifically designed biopanning schemes can now yield scFvs possessing specific recognition capabilities. A rational mutation strategy further enhances the affinity of scFv, and allows it to reach a level that cannot be achieved by immunization. Finally, appropriate prokaryotic expression measures ensure stable and efficient production of scFv. Therefore, when developing excellent scFvs, it is necessary to focus on three key aspects of this process that include screening, mutation, and expression. In this review, we analyze in detail the preparation and affinity improvement process for scFv and provide insights into the research progress and development trend of scFv-based immunoassay methods for monitoring small-molecule harmful substances.

Wood-based multi-layered porous imprinted membranes with dual-metal composite nanostructures: Synergistic promotion of rebinding capacity, selectivity and permeability

Herein the Ag&TiO2-based propranolol-imprinted layers had been prepared onto basswood membrane surface based on a polydopamine (PDA)-induced self-polymerization/composite strategy. The as-prepared PDA-induced Ag&TiO2-based molecularly imprinted nanocomposite membranes (PAT-MIMs) showed superb specific recognition capability and permselectivity toward the template molecule of propranolol. Thereinto, the natural curved channel of basswood could efficiently promote the diffusion process of propranolol and improve the contact efficiency with surface recognition sites. Describing the entire synthetic strategy, the as-prepared PDA-induced Ag&TiO2-modified basswood-based membrane structure had been regarded as a threefold sol-gel imprinted-initiated system, thus constructing the propranolol-imprinted sites with key properties of high selectivity and rapid specific-recognizability. Therefore, enhanced adsorption performance of 118.6 mg/g, fast adsorption rate (within 30 min) and high permselectivity rates (more than 3.0) had been successfully harvested, which clearly illustrated the successful preparation of high-intensitive recognizability and permeability toward propranolol from complex separation system.

PH-redox responsive cascade-targeted liposomes to intelligently deliver doxorubicin prodrugs and lonidamine for glioma

To synergistically treat glioma with a combination chemotherapy, we design and prepare novel cascade-targeted liposomes (Lip-TPGS) using glucose and triphenylphosphonium (TPP) as targeting moieties, which could intelligently deliver redox-sensitive doxorubicin (DOX) prodrugs (SDOX) and chemotherapeutic sensitizer lonidamine (LND). The pH-responsive ligand Chol-TPG modified by PEGylated glucose can overcome the blood-brain barrier and reach tumor cells. Combined with the modification of mitochondria targeting ligand (Chol-TPP), Lip-TPGS are endowed with pH-responsive charge regulation function and multi-stage targeting abilities. After triggered by the excessive glutathione in tumor cells, Lip-TPGS could sufficiently release the parent drugs DOX, which would significantly reduce side effects without compromising anti-glioma efficacy. Therefore, Lip-TPGS possess these characteristics: good pharmacokinetic behavior, superior brain targeting ability, specific tumor recognition and internalization capability, and strong endo/lysosome escaping and mitochondria targeting potential. Furthermore, Lip-TPGS exhibit significant advantages on anti-glioma by inhibiting proliferation, promoting apoptosis, inducing mitochondria dysfunction, inhibiting migration and invasion, prolonging the survival time, narrowing tumor areas, limiting lung metastasis, and reducing toxicity to normal organs. In summary, Lip-TPGS, with cascade targeting abilities from tissue/cell to organelle levels and highly controlled drug release properties, would become a promising drug delivery system for glioma treatment.

Development of magnetic molecularly imprinted polymers with double templates for the rapid and selective determination of carbamazepine and lamotrigine in serum.

A dual-template magnetic molecularly imprinted polymer (Dt-MMIP) with a specific recognition capability for carbamazepine (CBZ) and lamotrigine (LTG) was synthesized using methacrylic acid as a functional monomer, and ethylene glycol dimethylmethacrylate as a cross-linking agent. A magnetic non-molecularly imprinted polymer without templates (MNIP) was also prepared using the same procedure. The prepared polymers were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy and adsorption experiments. Results indicated that both Dt-MMIPs and MNIPs were microspherical nanoparticles, and the surface of the Dt-MMIP was rougher than that of the MNIP. In addition, the prepared Dt-MMIPs possessed a higher adsorption capacity and better selectivity for CBZ and LTG than the MNIPs. The maximum static adsorption capacities of Dt-MMIP for CBZ and LTG were 249.5 and 647.9 μg g−1, respectively, whereas those of MNIP were 75.8 and 379.8 μg g−1, respectively. The obtained Dt-MMIPs were applied as a magnetic solid-phase extraction sorbent for the rapid and selective extraction of CBZ and LTG in rat serum samples, and determination was performed by high-performance liquid chromatography with UV detection (HPLC-UV). The developed method of dispersive SPE based on Dt-MMIPs coupled to HPLC-UV has good rapidity and selectivity, and application prospects in serum.

DNA Aptamers Block the Receptor Binding Domain at the Spike Protein of SARS-CoV-2.

DNA aptamers are versatile molecular species obtained by the folding of short single-stranded nucleotide sequences, with highly specific recognition capabilities against proteins. Here we test the ability of DNA aptamers to interact with the spike (S-)protein of the SARS-CoV-2 viral capsid. The S-protein, a trimer made up of several subdomains, develops the crucial function of recognizing the ACE2 receptors on the surface of human cells, and subsequent fusioning of the virus membrane with the host cell membrane. In order to achieve this, the S1 domain of one protomer switches between a closed conformation, in which the binding site is inaccessible to the cell receptors, and an open conformation, in which ACE2 can bind, thereby initiating the entry process of the viral genetic material in the host cell. Here we show, by means of state-of-the-art molecular simulations, that small DNA aptamers experimentally identified can recognize the S-protein of SARS-CoV-2, and characterize the details of the binding process. We find that their interaction with different subdomains of the S-protein can effectively block, or at least considerably slow down the opening process of the S1 domain, thereby significantly reducing the probability of virus-cell binding. We provide evidence that, as a consequence, binding of the human ACE2 receptor may be crucially affected under such conditions. Given the facility and low cost of fabrication of specific aptamers, the present findings could open the way to both an innovative viral screening technique with sub-nanomolar sensitivity, and to an effective and low impact curative strategy.

All-Dielectric Metasurface for Sensing Microcystin-LR

Sensing Microcystin-LR (MC-LR) is an important issue for environmental monitoring, as the MC-LR is a common toxic pollutant found in freshwater bodies. The demand for sensitive detection method of MC-LR at low concentrations can be addressed by metasurface-based sensors, which are feasible and highly efficient. Here, we demonstrate an all-dielectric metasurface for sensing MC-LR. Its working principle is based on quasi-bound states in the continuum mode (QBIC), and it manifests a high-quality factor and high sensitivity. The dielectric metasurface can detect a small change in the refractive index of the surrounding environment with a quality factor of ~170 and a sensitivity of ~788 nm/RIU. MC-LR can be specifically identified in mixed water with a concentration limit of as low as 0.002 μg/L by a specific recognition technique for combined antigen and antibody. Furthermore, the demonstrated detection of MC-LR can be extended to the identification and monitoring of other analytes, such as viruses, and the designed dielectric metasurface can serve as a monitor platform with high sensitivity and high specific recognition capability.

Function-Oriented Graphene Quantum Dots Probe for Single Cell in situ Sorting of Active Microorganisms in Environmental Samples.

Functional microorganisms play a vital role in removing environmental pollutants because of their diverse metabolic capability. Herein, a function-oriented fluorescence resonance energy transfer (FRET)-based graphene quantum dots (GQDs-M) probe was developed for the specific identification and accurate sorting of azo-degrading functional bacteria in the original location of environmental samples for large-scale culturing. First, nitrogen-doped GQDs (GQDs-N) were synthesized using a bottom-up strategy. Then, a GQDs-M probe was synthesized based on bonding FRET-based GQDs-N to an azo dye, methyl red, and the quenched fluorescence was recovered upon cleavage of the azo bond. Bioimaging confirmed the specific recognition capability of GQDs-M upon incubation with the target bacteria or environmental samples. It is suggested that the estimation of environmental functional microbial populations based on bioimaging will be a new method for rapid preliminary assessment of environmental pollution levels. In combination with a visual single-cell sorter, the target bacteria in the environmental samples could be intuitively screened at the single-cell level in 17 bacterial strains, including the positive control Shewanella decolorationis S12, and were isolated from environmental samples. All of these showed an azo degradation function, indicating the high accuracy of the single-cell sorting strategy using the GQDs-M. Furthermore, among the bacteria isolated, two strains of Bacillus pacificus and Bacillus wiedmannii showed double and triple degradation efficiency for methyl red compared to the positive control (strain S12). This strategy will have good application prospects for finding new species or high-activity species of specific functional bacteria.

Mechanism of selective gold adsorption on ion-imprinted chitosan resin modified by thiourea

Thiourea-modified chitosan-imprinted resin (IM-TUCS) and a corresponding nonimprinted resin (NIM-TUCS) were synthesized and characterized using adsorption experiments. The adsorption results showed that adsorption reached equilibrium within 4 h. The adsorption data were better fitted using the Langmuir model (R2>0.99), and the gold adsorption capacities of IM-TUCS and NIM-TUCS were 933.2 and 373.7 mg·g−1, respectively. The IM-TUCS adsorbent was more suitable for gold than other coexisting anions and cations. The possible mechanism underlying Au(Ⅲ) adsorption on IM-TUCS was further investigated using X-ray photoelectron spectroscopy (XPS), Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction analyses. The protonation of the amino group on the resin under low pH conditions promoted Au(Ⅲ) adsorption; O, N and S in the C‒OH, C˭S and C-NH2 groups contained in the IM-TUCS coordinated with Au(III) ions. The cross-linking of the imprinted resin provided holes that could hold Au(III), thus the imprinted resin supported more Au(III). The adsorption capacity of the IM-TUCS for Au(III) was significantly higher than that of the NIM-TUCS, which is attributed to the cross-linking of the imprinted resin. Moreover, the IM-TUCS showed specific recognition capabilities for Au(III). After elution with the eluent, IM-TUCS was reused for four cycles with a gold recovery rate of approximately 93%, revealing its high potential economic value.

Metal-Organic Gel-Modulated Synthesis of Hierarchically Porous Molecularly Imprinted Polymers for Efficient Removal of Sildenafil from Water.

Molecularly imprinted polymers (MIPs) with high specific recognition capability are promising in environmental remediation. However, traditional MIPs usually show poor specific binding affinity toward templates in pure aqueous medium, thus greatly limiting their practical applications in wastewater treatment. Herein, we proposed a facile and versatile method to synthesize a water-compatible hierarchically porous MIP (HP-MIP), in which a metal–organic gel (MOG) was formed by in situ assembly and acted as a removable structural modulator. Remarkably, the integration of the MOG modulator and template imprinting defects significantly improved the specific template binding affinity of HP-MIP in water. The adsorption behavior of HP-MIP fitted well with the heterogeneous Freundlich isotherm, suggesting that HP-MIP possessed greater site heterogeneity to sildenafil than HP-NIP, which confirmed the efficiency of HP-MIP for the removal of sildenafil from water. This approach provides an important pathway to prepare water-compatible porous MIP for efficient removal of highly toxic organic pollutants from wastewater.

Gold Nanostars Bioconjugation for Selective Targeting and SERS Detection of Biofluids.

Gold nanoparticles (AuNPs) show physicochemical and optical functionalities that are of great interest for spectroscopy-based detection techniques, and especially for surface enhanced Raman spectroscopy (SERS), which is capable of providing detailed information on the molecular content of analysed samples. Moreover, the introduction of different moieties combines the interesting plasmonic properties of the AuNPs with the specific and selective recognition capabilities of the antibodies (Ab) towards antigens. The conjugation of biomolecules to gold nanoparticles (AuNPs) has received considerable attention for analysis of liquid samples and in particular biological fluids (biofluids) in clinical diagnostic and therapeutic field. To date, gold nanostars (AuNSts) are gaining more and more attention as optimal enhancers for SERS signals due to the presence of sharp branches protruding from the core, providing a huge number of “hot spots”. To this end, we focused our attention on the design, optimization, and deep characterization of a bottom up-process for (i) AuNPs increasing stabilization in high ionic strength buffer, (ii) covalent conjugation with antibodies, while (iii) retaining the biofunctionality to specific tag analyte within the biofluids. In this work, a SERS-based substrate was developed for the recognition of a short fragment (HA) of the hemagglutinin protein, which is the major viral antigen inducing a neutralizing antibody response. The activity and specific targeting with high selectivity of the Ab-AuNPs was successfully tested in transfected neuroblastoma cells cultures. Then, SERS capabilities were assessed measuring Raman spectra of HA solution, thus opening interesting perspective for the development of novel versatile highly sensitive biofluids sensors.

Guanosine-rich aptamers@Cu2O nanoparticles: enhanced peroxidase activity and specific recognition capability at neutral pH.

The decoration of Cu2O nanoparticles with guanosine-rich aptamers can significantly enhance their peroxidase activity at neutral pH and endow them with specific recognition capabilities. Both the phosphate backbone and guanine of the aptamers contribute to the enhancement. The excellent enzyme-like properties of this Cu2O-aptamer system make it a versatile platform for the development of neutral pH biosensors.

Biomolecule-tailored assembly and morphology of gold nanoparticles for LSPR applications

Gold nanoparticles (GNPs) have attracted the particular attention of nanoscience and nanoengineering due to their unique localized surface plasmonic resonance (LSPR) derived optical, electronic and magnetic properties. These LSPR-based properties are highly dependent on the nanostructures of GNPs which could be precisely tuned via strategies of assembly and morphological regulation. Biomolecules (DNA, amino acids and peptides) serving as linking and capping agents are capable to program the assembling and tailor the morphological evolution of GNPs to achieve varying nanostructures and plasmonic properties as well as biofunctionalities due to their specific recognition capabilities and unique interface affinity with GNPs. Versatile applications based on tunable LSPR properties of plasmonic nanoprobes composed of GNPs and biomolecules, therefore, have been successfully demonstrated in various fields, including metal-enhanced fluorescence, dark-field microscopy imaging, colorimetric assay, plasmonic circular dichroism-based assay, surface-enhanced Raman scattering-based bioprobes and LSPR shift-based biosensing, etc. This review introduces the recent advances in the design, structural tuning, property characterization and biosensing and bioimaging applications of these plasmonic nanoprobes, with detailed examples in previous applications and prospective discussions for their developments in future.