Protein-Protein Binding Kinetics by Biolayer Interferometry.

The specific kinetics and thermodynamics of protein-protein interactions underlie the molecular mechanisms of cellular functions; hence the characterization of these interaction parameters is central to the quantitative understanding of physiological and pathological processes. Many methods have been developed to study protein-protein interactions, which differ in various features including the interaction detection principle, the sensitivity, whether the method operates in vivo, in vitro, or in silico, the temperature control, the use of labels, immobilization, the amount of sample required, the number of measurements that can be accomplished simultaneously, or the cost. Bio-Layer Interferometry (BLI) is a label-free biophysical method to measure the kinetics of protein-protein interactions. Label-free interaction assays are a broad family of methods that do not require protein modifications (other than immobilization) or labels such as fusions with fluorescent proteins or transactivating domains or chemical modifications like biotinylation or reaction with radionuclides. Besides BLI, other label-free techniques that are widely used for determining protein-protein interactions include surface plasmon resonance (SPR), thermophoresis, and isothermal titration calorimetry (ITC), among others.

Advancing the study of protein-G4 interactions in DNA repair: Insights from biolayer interferometry.

Biolayer interferometry (BLI) is a powerful tool that enables direct observations of protein-G4 interactions in real-time. In this article, we discuss the crucial aspects in conducting a BLI experiment by using the TAR DNA-binding protein (TDP43) and a G4 DNA formed by (GGGGCC)4 as a sample application. We also describe the necessary precautions in designing the DNA substrate and evaluating the signal contributions arising from nonspecific binding interactions. A comprehensive guide is included that details the necessary materials and reagents, experimental procedures, and data analysis methods for researchers who are interested in using BLI for similar studies. The insights provided in this article will allow researchers to harness the potential of BLI and unravel the complexities of protein-G4 interactions with precision and confidence.

Analysis and prediction of ground deformation in Yinxi Industrial Park based on time-series InSAR technology.

Monitoring ground deformation in industrial parks is of great importance for the economic development of urban areas. However, limited research has been conducted on the deformation mechanism in industrial parks, and there is a lack of integrated monitoring and prediction models. Therefore, this study proposes a comprehensive monitoring and prediction model for industrial parks, utilizing time-series Interferometry Synthetic Aperture Radar (InSAR) technology and the Whale Optimization Algorithm-Back Propagation (WOA-BP) neural network algorithm. Taking Yinxi Industrial Park in Baiyin District as a case study, we used 68 scenes of Sentinel-1A ascending and descending orbit data from June 2018 to April 2021. The Stanford Method for Persistent Scatterers-Permanent Scatterers (StaMPS-PS) and the Small Baseline Subsets-Interferometry Synthetic Aperture Radar (SBAS-InSAR) technologies were employed to obtain the surface deformation information of the park. The deformation information obtained by the two technologies was cross-validated in terms of temporal and spatial distribution, and the vertical and east-west deformation of the park was obtained by combining the ascending and descending orbit data. The results show that the deformation feature points in the line of sight (LOS) direction obtained by the two technologies have a high consistency in spatial distribution, using the ascending orbit data as an example. Additionally, the SBAS-InSAR technology was used to obtain the east-west and vertical deformation results of the park after merging the ascending and descending orbit data for the same period. It was found that the park is mainly affected by vertical deformation, with a maximum subsidence rate of 14.67 mm/yr. The subsidence areas correspond to the deformation positions observed in field survey photos. Based on the ascending orbit deformation data, the two technologies were validated with 585 points of the same latitude and longitude, and the coefficient of determination R2 was found to be 0.82, with a root mean square error (RMSE) of 2.20 mm/a. The deformation rates were also highly consistent. Due to the 47% increase in the number of sampling points provided by the StaMPS-PS technique compared to the SBAS-InSAR technique, the former was found to be more applicable in the industrial park. Based on the ground deformation mechanism in the park, we combined the StaMPS-PS technique with the WOA-BP neural network to construct a deformation zone prediction model. We conducted predictive studies on the deformation zones of buildings and roads within the park, and the results showed that the WOA-optimized BP neural network achieved higher accuracy and lower overall error compared to the unoptimized network. Finally, we analyzed and discussed the geological conditions and inducing factors of ground deformation in the park, providing a reference for a better understanding of the deformation mechanism and early warning of disasters in the industrial park.

Comparative analysis of aptamers binding to SARS-CoV-2 N protein using capillary electrophoresis and bio-layer interferometry.

Aptamers are increasingly employed in SARS-CoV-2 theragnostics in recent years. Characterization of aptamers, testing affinity and kinetic parameters (e.g., equilibrium dissociation constant (KD), kon, and koff), can be done by several methods and influenced by many factors. This study aims to characterize the binding of aptamers to SARS-CoV-2 nucleocapsid (N) protein using capillary electrophoresis (CE) and bio-layer interferometry (BLI). These two analytical methods differ by how the aptamer binds to its target protein once the aptamer, as a capture ligand, is partitioned in solution (CE) or immobilized on the biosensor (BLI). With CE, the KD values of the N-binding aptamers (tNSP1, tNSP2, and tNSP3) were determined to be 18 ± 4 nM, 45 ± 11 nM, and 32 ± 7 nM, respectively, while the KD measurements by BLI yielded 4.8 ± 0.6, 4.5 ± 0.5, and 2.9 ± 0.3 nM, respectively. CE results showed a higher KD across all aptamers tested. The differences in the steric hindrance and confirmational structures of the aptamers immobilized on the BLI biosensors versus those suspended in the CE sample solution affect the molecular interactions between aptamers and the target proteins. Moreover, the buffer composition including pH and ionic strength can influence the stability of aptamer structures, or aptamer-protein complexes. All these variables affect the binding and calculated KD. In this sense, a KD value alone is not sufficient to make comparisons between aptamers; instead, the entire experimental setup should also be considered. This is particularly important when implementing aptamers in different bioanalytical systems.

Real-time tracking of the adsorption of bovine serum albumin on lipid layer and its effect on lipolysis by optical interferometry.

The model proteins bovine serum albumin (BSA) and lipid layer were used to study the effect of proteins on lipolysis. A lipid layer with an interference effect was constructed by loading the triolein into the silica colloidal crystal (SCC) film. The ordered porous layer interferometry (OPLI) system was used to track the changes in lipid layer mass caused by lipase hydrolysis to achieve real-time lipolysis detection. The real-time tracking of the adsorption of BSA on the lipid layer by converting the migration of interference fringes caused by the change of the lipid layer into the optical thickness change (ΔOT). The effect of BSA on the early and late stages of lipolysis was studied, and lipases containing 5 mg/mL BSA degraded the lipid layer 3.4 times faster than lipases containing 0.1 mg/mL BSA in the later stages. This study deepens the understanding of protein-lipid interactions in complex digestive environments.

An interferometric-based tensile tester to resolve damage events within reconstituted multi-filaments collagen bundles.

Understanding how mechanical damage propagates in load-bearing tissues such as skin, tendons and ligaments, is key to developing regenerative medicine solutions for when these tissues fail. For collagenous tissues in particular, damage is typically assessed after mechanical testing using a broad range of microscopy techniques because standard tensile testing systems do not have the time and force sensitivity to resolve mechanical damage events. Here we introduce an interferometric detection scheme to measure the displacement of a cantilever with a resolution of 0.03% of full scale at a sampling rate of 5000 samples/s. The system is validated using collagen fibers engineered to mimic mammalian tendons. The system can detect sudden decrease in force due to slippage between collagen filaments, one to five microns in diameter, within a fiber in air. It can also detect yield events associated with local collagen unfolding or sliding within collagen fibrils within a fiber in liquid. This is opening the road to the sub-failure study of damage propagation within a broad range of hierarchical biomaterials.

Optical fiber biosensors toward in vivo detection.

This review takes stock of the various optical fiber-based biosensors that could be used for in vivo applications. We discuss the characteristics that biosensors must have to be suitable for such applications and the corresponding transduction modes. In particular, we focus on optical fiber biosensors based on fluorescence, evanescent wave, plasmonics, interferometry, and Raman phenomenon. The operational principles, implemented solutions, and performances are described and debated. The different sensing configurations, such as the side- and tip-based fiber biosensors, are illustrated, and their adaptation for in vivo measurements is discussed. The required implementation of multiplexed biosensing on optical fibers is shown. In particular, the use of multi-fiber assemblies, one of the most optimal configurations for multiplexed detection, is discussed. Different possibilities for multiple localized functionalizations on optical fibers are presented. A final section is devoted to the practical in vivo use of fiber-based biosensors, covering regulatory, sterilization, and packaging aspects. Finally, the trends and required improvements in this promising and emerging field are analyzed and discussed.

Interaction studies between human papillomavirus virus-like particles and laminin 332 by affinity capillary electrophoresis assisted by bio-layer interferometry.

Human papillomavirus (HPV) interacts, in vitro, with laminin 332 (LN332), a key component of the extracellular matrix. In this study, we performed bio-layer interferometry (BLI) and affinity capillary electrophoresis (ACE) to investigate the binding properties of this interaction. Virus-like particles (VLPs), composed of the HPV16 L1 major capsid protein, were used as HPV model and LN332 as the VLPs binding partner. Using BLI, we quantitatively determined the kinetics of the interaction, via the measurement of VLP binding and release from LN332 immobilized onto the surface of aminopropylsilane biosensors. We found an averaged kon of 1.74 x 104 M-1s-1 and an averaged koff of 1.50 x 10-4 s-1. Furthermore, an ACE method was developed to study the interaction under physiological conditions, where the interactants are moving freely in solution, without any fluorescence labeling. Specifically, a constant amount of HPV16-VLPs was preincubated with increasing LN332 concentrations and then the samples were injected in the capillary electrophoresis instrument. A shift in the migration time of the HPV16-VLP/LN332 complexes, carrying an increasing number of LN332 molecules bound per VLP, was observed. The mobility of the complexes was found to decrease with increasing LN332 concentrations in the sample. It was used to quantify stability constant. From BLI and ACE approaches, we reported an apparent equilibrium dissociation constant in the nanomolar range (8.89 nM and 17.7 nM, respectively) for the complex between HPV16-VLPs and LN332.

Analyzer-free hard x-ray interferometry.

Objective. To enable practical interferometry-based phase contrast CT using standard incoherent x-ray sources, we propose an imaging system where the analyzer grating is replaced by a high-resolution detector. Since there is no need to perform multiple exposures (with the analyzer grating at different positions) at each scan angle, this scheme is compatible with continuous-rotation CT apparatus, and has the potential to reduce patient radiation dose and patient motion artifacts.Approach. Grating-based x-ray interferometry is a well-studied technique for imaging soft tissues and highly scattering objects embedded in such tissues. In addition to the traditional x-ray absorption-based image, this technique allows reconstruction of the object phase and small-angle scattering information. When using conventional incoherent, polychromatic, hard x-ray tubes as sources, three gratings are usually employed. To sufficiently resolve the pattern generated in these interferometers with contemporary x-ray detectors, an analyzer grating is used, and consequently multiple images need to be acquired for each view angle. This adds complexity to the imaging system, slows image acquisition and thus increases sensitivity to patient motion, and is not dose efficient. By simulating image formation based on wave propagation, and proposing a novel phase retrieval algorithm based on a virtual grating, we assess the potential of a analyzer-grating-free system to overcome these limitations.Main results. We demonstrate that the removal of the analyzer-grating can produce equal image contrast-to-noise ratio at reduced dose (by a factor of 5), without prolonging scan duration.Significance.By demonstrating that an analyzer-free CT system, in conjuction with an efficient phase retrieval algorithm, can overcome the prohibitive dose and workflow penalties associated grating-stepping, an alternative path towards realizing clinical inteferometric CT appears possible.

Advanced Interferometry with 3-D Structured Illumination Reveals the Surface Fine Structure of Complex Biospecimens.

Interference reflection microscopy (IRM) is a powerful, label-free technique to visualize the surface structure of biospecimens. However, stray light outside a focal plane obscures the surface fine structures beyond the diffraction limit (dxy ≈ 200 nm). Here, we developed an advanced interferometry approach to visualize the surface fine structure of complex biospecimens, ranging from protein assemblies to single cells. Compared to 2-D, our unique 3-D structure illumination introduced to IRM enabled successful visualization of fine structures and the dynamics of protein crystal growth under lateral (dx-y ≈ 110 nm) and axial (dx-z ≤ 5 nm) resolutions and dynamical adhesion of microtubule fiber networks with lateral resolution (dx-y ≈ 120 nm), 10 times greater than unstructured IRM (dx-y ≈ 1000 nm). Simultaneous reflection/fluorescence imaging provides new physical fingerprints for studying complex biospecimens and biological processes such as myogenic differentiation and highlights the potential use of advanced interferometry to study key nanostructures of complex biospecimens.

Monitoring land subsidence in the Peshawar District, Pakistan, with a multi-track PS-InSAR technique.

Peshawar is one of the most densely populated cities of Pakistan with high urbanization rate. The city overexploits groundwater resources for household and commercial usage which has caused land subsidence. Land subsidence has long been an issue in Peshawar due to insufficient groundwater removal. In this research, we employ the persistent scatterer interferometry synthetic aperture radar (PS-InSAR) technique with Sentinel-1 imaging data to observe the yearly land subsidence and generate accumulative time-series maps for the years (2018 to 2020) using the SAR PROcessing tool (SARPROZ). The PS-InSAR findings from two contiguous paths are combined by considering the variance over the overlapping area. The subsidence rates in the Peshawar are from -59 to 17 mm/yr. The results show that subsidence is -28.48 mm/yr in 2018, the subsidence reached -49.02 mm/yr in 2019, while in 2020, the subsidence reached -49.90 mm/yr. The findings indicate a notable rise in land subsidence between the years 2018 and 2020. Subsidence is predicted in the research region primarily due to excessive groundwater removal and soil consolidation induced by surficial loads. The correlation of land subsidence observations with groundwater levels and precipitation data revealed some relationships. Overall, the proposed method efficiently monitors, maps, and detects subsidence-prone areas. The utilization of land subsidence maps will enhance the efficiency of urban planning, construction of surface infrastructure, and the management of risks associated with subsidence.

Measurement of enzyme activity of insoluble substrates based on ordered porous layer interferometry and the application in evaluation of thrombolytic drugs.

The development of innovative methods for real-time surveillance of enzymatic activity determination processes is essential, particularly for insoluble substrate enzymatic assessments. In this work, a novel method for enzymatic activity determination was devised by assembling a 190 nm silica colloidal crystal (SCC) film onto a glass slide, coupled with Ordered Porous Layer Interferometry (OPLI) technology. By fixing the substrate of the enzyme on the surface of the silica sphere, a solid-liquid interface can be formed for monitoring enzymatic activity. The enzymatic activity is gauged by the change in the SCC film's thickness caused by the digestion of the loaded substrate. The procedure of chymotrypsin-mediated casein digestion was documented in real time, facilitating the examination of chymotrypsin's activity and kinetics. The newly-developed enzymatic activity determination method demonstrated exceptional sensitivity towards chymotrypsin activity, with a linear range spanning 0.0505-2.02 units per mg. Additionally, the method was extended to the assessment of fibrinolysis enzyme activity and kinetic analysis, yielding promising results. Therefore, this technique can serve as a real-time, user-friendly, cost-effective novel approach for enzymatic activity determination, providing fresh perspectives for enzymatic activity determination studies.

Advanced control strategies for continuous capture of monoclonal antibodies based upon biolayer interferometry.

The semi and fully continuous production of monoclonal antibodies (mAbs) has been gaining traction as a lower cost, and efficient production of mAbs to broaden patient access. To be truly flexible and adaptive to process demands, the industry has lacked sufficient advanced control strategies. The variation of the upstream product concentration typically cannot be handled by the downstream capture step, which is configured for a constant feed concentration and fixed binding capacity. This inflexibility leads to losses of efficiency and product yield. This study shows that these challenges can be overcome by a novel advanced control strategy concept that includes dynamic control throughout a perfusion bioreactor, with cell retention by alternating tangential flow, integrated with simulated moving bed (SMB) multi-column chromatography. The automation workflow and advanced control strategy were implemented through the use of a visual programming development environment. This enabled dynamic flow control across the upstream and downstream process integrated with a dynamic column loading of the SMB. A sensor prototype, based on continuous biolayer interferometry measurements was applied to detect mAb breakthrough within the last column flow-through to manage column switching. This novel approach provided higher specificity and lower background signal compared to commonly used spectroscopy methods, resulting in an optimized resin utilization while simultaneously avoiding product loss. The dynamic loading was found to provide a twofold increase of the mAb concentration in the eluate compared to a conservative approach with a predefined recipe with similar impurity removal. This concept shows that advanced control strategies can lead to significant process efficiency and yield improvement.

Reliability and Agreement of Keratometry Measurements Obtained With Eye Surface Profilometry and Partial Coherence Interferometry.

PURPOSE: The aim of this study was to assess the repeatability of keratometry parameters obtained using the Eye Surface Profiler (ESP) system and their agreement with the IOL Master 500 device. METHODS: Seventy-one eyes of 71 healthy participants were evaluated. Three repeated measurements were performed using the ESP system. Simulated keratometry in the flat (SimKf) and steep (SimKs) meridians, astigmatism, and axis were obtained. The same parameters were measured using the IOL Master 500 device. The J0 and J45 vector components of the astigmatism were calculated. The intrasession repeatability was analyzed using within-subject SD (Sw) and intraclass correlation coefficient (ICC). Agreement was assessed using paired statistical tests and the Bland-Altman method. RESULTS: The Sw was 0.07 mm, 0.04 mm, 0.51 D, 0.33 D, and 0.22 D, and the ICC was 0.96, 0.98, 0.74, 0.61, and 0.55 for SimKf, SimKs, astigmatism, J0, and J45, respectively. The mean difference and limits of agreement when comparing the ESP system with the IOL Master 500 device were 0.37 mm (0.08/0.66) for SimKf ( P < 0.001), 0.18 mm (0.00/0.35) for SimKs ( P < 0.001), -0.93 D (-2.42/0.56) for astigmatism ( P < 0.001), 0.51 D (-0.22/1.24) for J0 ( P < 0.001), and 0.06 D (-0.48/0.60) for J45 ( P = 0.09). CONCLUSIONS: The ESP system provides consistent values for simulated keratometry, showing moderate consistency for astigmatism parameters. Contact lens practitioners should be aware that the ESP system and IOL Master 500 device provide different simulated keratometry from a clinically viewpoint.

Highly-efficient selection of aptamers for detecting various HPV subtypes in clinical samples.

Nucleic acid aptamers are of great potentials in diagnostic and therapeutic applications because of their unique molecular recognition capabilities. However, satisfactory aptamers with high affinity and specificity are still in short supply. Herein, we have developed new selection methods allowing the free interactions between the targets and potential aptamers in solution. In our selection system, the protein targets (biotinylated randomly or site-specifically) were first incubated with the random DNA library, followed by the pull-down with the streptavidin magnetic beads or biolayer-interferometry (BLI) sensors. By comparing the two biotinylation strategies (random or site-specific) and two states of the targets (free or immobilized), we have found that the combination of the site-specific biotinylation and free-target strategies was most successful. Based on these highly-efficient selection strategies, HPV L1 aptamers were obtained. By designing the sandwich aptasensor assisted with RCA and CRISPR/Cas12a, we have diagnosed various HPV subtypes in clinical samples, such as easily-collected urine samples. In summary, our new strategy can allow efficient selection of aptamers with high affinity and specificity for clinical applications.

Distributed interferometric fiber tip biosensors for a multi-channel and label-free biomolecular interaction analysis.

This paper describes fabrication and implementation of distributed optical fiber tip biosensor probes for simultaneously measuring label-free biomolecular interactions at multiple locations. Biosensor probes at the tip of a single-mode fiber are Fabry-Perot etalons that are functionalized with a capture layer for a specific biomolecule. A coherence multiplexing method is implemented to separate data collected from distributed biosensors in a single data stream. Multiplexing is achieved by using fiber tip biosensors of varying etalon lengths with the same or different capture layers for each biosensing channel. Experiments demonstrating simultaneous multi-channel recording of protein-to-protein interaction sensorgrams with fiber tip biosensor probes are presented.

Decoding diffuse light scattering dynamics in layered tissues: path length versus fluctuation time scale.

Dynamic multiple light scattering (DMLS) has found numerous applications, including soft matter physics and biomedical optics. Yet biological tissues may have complex internal geometries, presenting a challenge for noninvasive measurements. Deciphering laminar dynamics is crucial to accurately interpret tissue or organ physiology. Seminal DMLS work noted that one can probe deeper layers indirectly by analyzing light fluctuations on shorter time scales. Recent technologies have enabled probing deeper layers directly by analyzing fluctuations at longer path lengths. The following question arises: are the indirect and direct approaches synergistic or redundant? Here, by adding an optical switch to path-length-filtered interferometric diffusing wave spectroscopy, we experimentally address this question in the context of a forearm occlusion study. We find that both approaches afford better distinction of light scattering dynamics in layered tissues than either approach alone. This motivates further development of methods that integrate both decorrelation time scale and light path length to probe layered tissues.

MAxSIM: multi-angle-crossing structured illumination microscopy with height-controlled mirror for 3D topological mapping of live cells.

Mapping 3D plasma membrane topology in live cells can bring unprecedented insights into cell biology. Widefield-based super-resolution methods such as 3D-structured illumination microscopy (3D-SIM) can achieve twice the axial ( ~ 300 nm) and lateral ( ~ 100 nm) resolution of widefield microscopy in real time in live cells. However, twice-resolution enhancement cannot sufficiently visualize nanoscale fine structures of the plasma membrane. Axial interferometry methods including fluorescence light interference contrast microscopy and its derivatives (e.g., scanning angle interference microscopy) can determine nanoscale axial locations of proteins on and near the plasma membrane. Thus, by combining super-resolution lateral imaging of 2D-SIM with axial interferometry, we developed multi-angle-crossing structured illumination microscopy (MAxSIM) to generate multiple incident angles by fast, optoelectronic creation of diffraction patterns. Axial localization accuracy can be enhanced by placing cells on a bottom glass substrate, locating a custom height-controlled mirror (HCM) at a fixed axial position above the glass substrate, and optimizing the height reconstruction algorithm for noisy experimental data. The HCM also enables imaging of both the apical and basal surfaces of a cell. MAxSIM with HCM offers high-fidelity nanoscale 3D topological mapping of cell plasma membranes with near-real-time ( ~ 0.5 Hz) imaging of live cells and 3D single-molecule tracking.

Multi-temporal SAR Interferometry (MTInSAR)-based study of surface subsidence and its impact on Krishna Godavari (KG) basin in India: a support vector approach.

The surface subsidence in the Krishna Godavari (KG) basin in India has increased with the discovery of crude oil and natural gas reserves since 1983. With private players coming up to bag the exploration and refining contracts, there must be timely monitoring of the surface subsidence of the region so that remedial measures for the resettlement of the populations can be taken promptly. Regular monitoring is necessary since the region is fertile and any seawater ingress results in the loss of valuable cultivable land. Multi-temporal SAR Interferometry (MTInSAR) technique has been applied successfully all over the world for the study and regular monitoring of land surface subsidence scenarios. This study utilizes data from Sentinel-1 C-band SAR sensor for MTInSAR-based surface subsidence and RADAR Vegetation Index (RVI)-based vegetation loss for the same season estimation between 2017 and 2022 for the KG basin region. It is inferred from the study that the region has shown surface subsidence of 80 mm/year between April 2020 and June 2022. This study uses support vector regressor (SVR) to predict the loss in forest cover in terms of RVI using MTInSAR-based surface subsidence, VH, and VV backscatter as parameters. It is observed that the SVR gave R2-statistics of 0.89 and 0.873 in the training and testing phases with a mean absolute error (MAE) and root mean squared error (RMSE) of 0.08 and 0.02, respectively. It is also observed that the region showed a loss of 3.21 km2 of cultivable land between 2020 and 2022.

Tapered Fiber Bioprobe Based on U-Shaped Fiber Transmission for Immunoassay.

In this paper, a tapered fiber bioprobe based on Mach-Zehnder interference (MZI) is proposed. To retain the highly sensitive straight-tapered fiber MZI sensing structure, we designed a U-shaped transmission fiber structure for the collection of optical sensing signals to achieve a miniature-insert-probe design. The spectrum responses from the conventional straight-tapered fiber MZI sensor and our proposed sensor were compared and analyzed, and experimental results showed that our proposed sensor not only has the same sensing capability as the straight-tapered fiber sensor, but also has the advantages of being flexible, convenient, and less liquid-consuming, which are attributed to the inserted probe design. The tapered fiber bioprobe obtained a sensitivity of 1611.27 nm/RIU in the refractive index detection range of 1.3326-1.3414. Finally, immunoassays for different concentrations of human immunoglobulin G were achieved with the tapered fiber bioprobe through surface functionalization, and the detection limit was 45 ng/mL. Our tapered fiber bioprobe has the insert-probe advantages of simpleness, convenience, and fast operation. Simultaneously, it is low-cost, highly sensitive, and has a low detection limit, which means it has potential applications in immunoassays and early medical diagnosis.