D-dimer is a soluble fibrin degradation product, and usually serves as a biomarker for screening and diagnosis of venous thromboembolism (VTE). Immunoturbidimetry is the most commonly used detection method in clinical practice, but it cannot achieve point-of-care testing (POCT) and on-site detection. In this study, two lateral flow immunochromatographic assay (LFIA), colloidal gold lateral flow immunoassay (CG-LFIA) and time-resolved fluorescence microsphere lateral flow immunoassay (TRFM-LFIA), were established for rapid qualification and accurate quantification of D-dimer in human plasma. Both analytical methods were rapid, simple, and capable of on-site monitoring without complex sample pretreatment. The CG-LFIA achieved rapid qualitative assessment based on the visible color intensity of the strips and on-site quantitative detection through strip image scanning. Compared with CG-LFIA, the TRFM-LFIA can provide a wider dynamic range and superior sensitivity for D-dimer detection, although it does not allow for rapid qualification. After systematic optimization and rigorous methodological validation, the detection limit of CG-LFIA was 40 ng/mL with a dynamic range of 40-1280 ng/mL, while TRFM-LFIA had a lower detection limit of 6.1 ng/mL and a wider dynamic range of 10-2000 ng/mL. Both methods demonstrated excellent consistency in detecting D-dimer in human plasma in comparison with immunoturbidimetry. Therefore, CG-LFIA is more suitable for rapid on-site screening of clinical thrombosis patients preliminarily, while TRFM-LFIA is more appropriate for rapid and accurate POCT quantification of Ddimer in human plasma. Both methods will provide more convenient testing services for clinical thrombus screening and thrombus progression assessment.

Surface plasmon resonance (SPR) is a sensitive, label-free method for measuring biomolecular interactions, widely used in drug discovery, including fishing active components from traditional Chinese medicine (TCM). However, classical SPR systems encounter limitations when dealing with the complexity of TCM, specifically regarding high-throughput processing and distinguishing substances with different binding kinetics. To overcome these challenges, we constructed an upgraded fluidic system (UFS) on the SPR platform, integrating a customized ligand fishing sensor chip, an optimized sample flow path, and a selectable recovery mode. By screening for tumor necrosis factor receptor 1 (TNFR1) ligands, we identified esculetin as a direct binding component. In SH-SY5Y cells, esculetin targets TNFR1, reducing endogenous ligand binding and decreasing caspase-3 activity, thereby inhibiting apoptosis. The UFS system significantly enhances screening efficiency for complex natural compounds. This study provides the first evidence that esculetin directly interacts with TNFR1 via a novel anti-apoptotic mechanism, complementing its known anti-inflammatory effects and offering a valuable tool for TCM research.

This study aims to develop an efficient and accurate method for detecting human serum albumin (HSA) in urine using lateral flow immunochromatography analysis (LFIA) and a smartphone-based application (App). First, a LFIA test strip using colloidal gold as the labeling material was developed and optimized. A detecting accessory was designed and produced, including a strip kit that can package the LFIA test strip, and a test cartridge that can mount the smartphone on top of it. Then an App was constructed to automatically identify the concentration of HSA on the test strip using photos taken by smartphone. The detection limit of HSA by the proposed platform is 2 µg/mL, and the quantitative range is 5 200 µg/mL. The coefficient of variation is less than 18.3

Surface plasmon resonance (SPR) is a sensitive, label-free method for measuring biomolecular interactions, widely used in drug discovery, including fishing active components from traditional Chinese medicine (TCM). However, classical SPR systems encounter limitations when dealing with the complexity of TCM, specifically regarding high-throughput processing and distinguishing substances with different binding kinetics. To overcome these challenges, we constructed an upgraded fluidic system (UFS) on the SPR platform, integrating a customized ligand fishing sensor chip, an optimized sample flow path, and a selectable recovery mode. By screening for tumor necrosis factor receptor 1 (TNFR1) ligands, we identified esculetin as a direct binding component. In SH-SY5Y cells, esculetin targets TNFR1, reducing endogenous ligand binding and decreasing caspase-3 activity, thereby inhibiting apoptosis. The UFS system significantly enhances screening efficiency for complex natural compounds. This study provides the first evidence that esculetin directly interacts with TNFR1 via a novel anti-apoptotic mechanism, complementing its known anti-inflammatory effects and offering a valuable tool for TCM research.

Surface plasmon resonance (SPR) biosensor has emerged as a transformative tool in high-throughput drug screening and label-free analysis of biomolecular interactions. However, a critical limitation of SPR lies in its stringent requirement for highly purified proteins to ensure reliable quantification of binding affinities and kinetic parameters. In order to address the limitations, the lentiviral particle and styrene-maleic acid polymer have been previously developed to extract and stabilize transmembrane proteins (TMs) indirectly and thus to detect ligands interaction with TMs by SPR biosensor. The present study proposes a high-throughput SPR-based drug screening system that utilizes cell-free protein synthesis (CFPS) to achieve in situ purification and immobilization of TMs on SPR biosensors. First, C−X−C chemokine receptor 4 (CXCR4) protein with His-tag was prepared by CFPS. Then, two types of nickel-nitrilotriacetic acid (Ni-NTA) biosensors, modified with carboxymethylated dextran (CMD) and coated with polycarboxylate hydrogel coating (HC) matrix, were compared with classical carboxymethylated dextran 5 (CM5) biosensor in order to determine the optimal strategy for coupling the CXCR4-CFPS protein. The CMD-NTA/SPR biosensor was next applied to screen for CXCR4 ligands from 96 natural products. Finally, glycyrrhizic acid and ginsenoside Re were proved to be function of CXCR4 inhibitors by affinity test, molecular docking, and cell migration assay. The combination of CFPS with SPR technology facilitates in-situ purification and immobilization of target proteins in a single step, thereby significantly enhancing the efficiency of SPR assay procedures. The system has broad applicability for targeting various challenging TMs and provides potential candidates for subsequent drug development.

The application of Bio-Layer Interferometry (BLI) is contingent upon the immobilization of highly purified target proteins onto the sensor. The cumbersome and time-consuming nature of traditional protein expression and purification processes restricts the application of BLI in high-throughput screening of traditional Chinese medicine (TCM). This study aims to develop a rapid and efficient BLI-based platform for screening bioactive components in TCM. An integrated platform combining cell-free protein synthesis (CFPS), BLI, and ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS) was established for efficient TCM bioactive compound discovery. Functional C-X-C chemokine receptor 4 (CXCR4) was synthesized in vitro using a CFPS system, which were then validated by surface plasmon resonance (SPR) and western blotting. Immobilized CXCR4 on NTA biosensors enabled BLI-based high-throughput screening of TCM extracts, followed by target-specific compound recovery and characterized via mass spectrometry. Three bioactive TCM constituents were successfully fished and identified as coptisine, ligustilide, and senkyunolide A. All of them exhibited negligible cytotoxicity at concentrations ranging from 6.25 to 100 μM). Furthermore, ligustilide and senkyunolide A demonstrated certain affinity for CXCR4 with KD of 69.86 μM and 14.7 μM, respectively, and significantly inhibited cell migration. This study is the first identification of ligustilide and senkyunolide A as functional ligands of CXCR4. The established CFPS-BLI-UHPLC-QTOF/MS platform enables efficient discovery of low-toxicity, high-affinity CXCR4-targeting therapeutics from TCM.

Ponkan Chenpi (PC), derived from the dried and aged peel of ripe Ponkan citrus, is a traditional product with both medicinal and edible value. In this study, an untargeted metabolomics approach utilizing UPLC-QTOF-MS/MS was employed to characterize the metabolite profiles of PCs aged for 1, 3, 5, and 8 years (PC1, PC3, PC5, and PC8). A total of 1455 metabolites were characterized, with flavonoids exhibiting relatively high quantity and abundance. Clear differences in both metabolite profiles and metabolic pathways were observed among PCs of different aging periods. Furthermore, flavonoid extracts were prepared from the PCs, with the content of flavonoid compounds determined by HPLC as the response value. The optimal extraction parameters, determined by single-factor analysis and response surface methodology, were 65 % ethanol concentration, 50 min ultrasonic time, 65 °C ultrasonic temperature, and a 1:65 solid-liquid ratio. Subsequent free radical scavenging assays demonstrated that the PC5 extract exhibited superior antioxidant activity. Correlation analysis indicated that flavonoid monomers—hesperidin, nobiletin, tangeretin, and 5-demethylnobiletin were significantly correlated with antioxidant activity. Network pharmacology and molecular docking analyses revealed that proteins such as PPARγ, COX-2, and ALB were the core targets through which these flavonoids exert antioxidant effects. Finally, the in vivo efficacy of PC5was verified in a D-gal-induced oxidative damage mouse model, which demonstrated that PC5 extract significantly improve the hepatic oxidative stress state and alleviate tissue damage in mice. This work provides a theoretical foundation for the quality evaluation and high-value utilization of PC.

Sini Decoction (SNT) is a traditional formula recognized for its efficacy in warming the spleen and stomach and dispersing cold. However, elucidating the mechanism of action of SNT remains challenging due to its complex multiple components. This study utilized a synergistic approach combining two-dimensional fluorescence difference in gel electrophoresis (2D-DIGE)-based drug affinity responsive target stability (DARTS) with label-free quantitative proteomics techniques to identify the direct and indirect protein targets of SNT in myocardial infarction. The analysis identified 590 proteins, with 30 proteins showing significant upregulation and 51 proteins showing downregulation when comparing the SNT group with the model group. Through the integration of 2D-DIGE DARTS with proteomics data and pharmacological assessments, the findings indicate that protein disulfide-isomerase A3 (PDIA3) may serve as a potential protein target through which SNT provides protective effects on myocardial cells during myocardial infarction.

Surface plasmon resonance (SPR) biosensors have been applied in various fields with the advantages of being label-free, having high specificity, having high sensitivity, and providing real-time monitoring. With the gradual improvement of SPR technology, SPR biosensors have been used for the detection of macromolecules such as proteins, peptides, and nucleic acids. Antibodies are generally used as the recognition component of SPR biosensors due to the high specificity of antibody–antigen binding. Recently, aptamers have become new choices instead of antibodies for their characteristic of high specificity with target molecules, high stability of chemical synthesis, convenience in storage, and ease of labeling. In this study, an aptamer-based SPR biosensor for chloramphenicol (CAP) detection was established through optimizing the conditions of CAP aptamer immobilization and analysis procedure, including biosensor type, signal enhancement, running buffer, sample diluent, and dissociation time. The results suggested that the optimal immobilization strategy of aptamers on the SPR biosensor was indirect immobilization based on the CM5 chip. The aptamer-based SPR biosensor had good specificity for CAP and could be used to detect CAP in real samples such as milk. Therefore, SPR biosensors have great application prospects in the food safety field, and aptamers deserve further study to improve the performance of the biosensor.

Background: Trichiosanthis Pericarpium (TP) is the dried ripe peel of Trichosanthes kirilowii Maxim., also known as gualoupi in Chinese, effectively clears heat and transforms phlegm. Traditional Chinese medicine (TCM) prescriptions that contain TP are widely used in clinical practice to treat respiratory diseases, including chronic obstructive pulmonary disease (COPD). However, the active ingredients of TP and the potential targets and mechanisms of action of TP against COPD have not been sufficiently investigated. Purpose: This study aimed to determine the active ingredients of TP and the potential targets and mechanisms of action of TP against COPD. Study design: The initial phase comprised the screening of potential active ingredients in TP, this was followed by the evaluation of their pharmacodynamic effects through both in vivo and in vitro experiments. Subsequently, network pharmacology and molecular docking were utilized to predict the key targets and associated pathways, which were later validated through animal-related experiments. Finally, the pharmacodynamic basis of TP interacting with the relevant target was identified using surface plasmon resonance (SPR). Methods: The potential active ingredients of TP were predicted by serum chemical composition analysis. The pharmacodynamic effect of Total Flavonoids of Trichiosanthis Pericarpium (TPTF) against COPD was demonstrated by in vivo and in vitro experiments. The targets and pathways of TPTF for COPD were predicted using network pharmacology and confirmed preliminarily by molecular docking techniques. The critical targets and pathways of TPTF against COPD were validated by Western blot and SPR. The active ingredients of TPTF were selected and identified through SPR. Results: The main active ingredients of TP are flavonoids, which are evaluated through serum chemical composition analysis. TPTF has been demonstrated to be effective in inhibiting inflammation and mucus hypersecretion in both in vivo and in vitro models of COPD. The targets of TPTF against COPD are focused on the EGFR/PI3K/AKT signaling pathway according to Network pharmacology, and the prediction was subsequently validated in the COPD mice. The flavonoids of TP that specifically target on EGFR include Luteolin-7-O-beta-D- glucoside, Quercetin-3-O-beta-rutinoside, and Apigenin-7-O-glucoside. Conclusion: This study demonstrates significant progress in understanding how the pharmacodynamic basis and mechanisms of TP improve COPD. The pharmacodynamic ingredients were identified as TPTF through predictions of serum chemical composition, experimental validation, and identification of SPR. The pharmacodynamic mechanisms were also derived from a comprehensive approach that combined network pharmacology, molecular docking predictions, experimental validation, and SPR identification. The innovative integration of different strategies has led to new findings that flavonoid glycosides, such as Luteolin-7-O-beta-D-glucoside, Quercetin-3-O-beta-rutinoside, and Apigenin-7-O-glucoside in TPTF, enhance the improvement of COPD by reducing inflammation and mucus hypersecretion associated with the EGFR/PI3K/AKT and EGFR/STAT3 signaling pathways.

Background: Cancer patients experience a high incidence of concomitant infections due to the effects of chemotherapy drugs and their suppressed immune function. Infection has become a major cause and an accelerating factor of cancer-related deaths. The combined use of anticancer drugs and antibiotics can produce adverse effects, necessitating the urgent search for dual-active drugs that are effective against both cancer and bacteria. Since tubulin has a homologous protein filamenting temperature-sensitive mutant Z (FtsZ) in bacteria, tubulin inhibitors have the potential to emerge as dual-active drugs against both cancer and bacteria. Methods: A comprehensive screening of a tubulin inhibitor library, encompassing 196 compounds, was conducted to evaluate their various activities. Results: Compounds 6, 23, 33, 56, 60, and 71 exhibited both anticancer and antibacterial activities in vitro, and 23, 33, 56, and 60 displayed varying degrees of FtsZ inhibitory activity. Particularly, compound 23 stood out as the most potent, exhibiting not only the strongest anticancer activity with IC50 values of 12, 20, and 10 nM against A549, MCF-7 and Hela cells, respectively, but also the most exceptional antibacterial activity with minimum inhibitory concentration (MIC) values of 8, 8, 64, and 32 μM against Staphylococcus aureus (S. aureus), Bacillus subtilis (B. subtilis), Escherichia coli (E. coli), and Pseudomonas aeruginosa (P. aeruginosa), respectively. Furthermore, compound 23 possessed the superior FtsZ inhibitory activity, facilitating polymerization. This was evident in the remarkably elongated cell morphology of Bacillus subtilis treated with compound 23. To gain a deeper understanding of the underlying mechanisms, molecular docking studies were conducted, revealing the interaction mode between compound 23 and both tubulin and FtsZ, further elucidating its multifaceted biological activities. Conclusions: The dual-active drugs obtained in this study provide a new solution to the problem of bacterial infection in cancer patients. The revealed FtsZ as the antibacterial target provides an important theoretical basis for further optimization of such drugs.

The development of programmed death 1 (PD-1) checkpoint/programmed death-ligand 1 (PD-L1) interaction inhibitors has opened a new front in the treatment of carcinoma. An increasing amount of research is devoted to small-molecule compounds that target this PD-1/PD-L1 interaction. In this article, we report the discovery of three new PD-1/PD-L1 inhibitors from Scutellaria baicalensis Georgi and Sophora flavescens Aiton herbal extracts, namely baicalin, maackiain, and oxysophocarpine, using a comprehensive ligand fishing system, which integrates a dual-target surface plasmon resonance biosensor and a magnetic beads method. These three compounds were also confirmed in the serum in vivo, validated by binding affinity evaluation, molecular docking, and competitive enzyme-linked immunosorbent assay (ELISA) assay to act upon the interface of PD-1/PD-L1. Thus, these three ingredients could be potential PD-1/PD-L1 inhibitors and may serve as hit compounds for immunotherapeutic drug discovery. These results also highlight the efficiency of the dual-target surface plasmon resonance (SPR) and magnetic beads ligand fishing system in drug screening for disease treatment.

Triple-negative breast cancer (TNBC) is limited in treatment options due to the absence of three receptors, and evidence suggests that TNBC is sensitive to ferroptosis. In this study, a series of genipin derivatives were synthesized through a hybridization strategy that integrated the structures of RSL3 and ML162. Among these derivatives, compound B23 demonstrated remarkable activity against the MDA-MB-231 cell line with an IC50 value of 40 nM, significantly outperforming genipin, RSL3 and ML162. Furthermore, B23 exhibited high selectivity for ferroptosis with a selectivity ratio of up to 108-fold. Further studies revealed that B23 induces ferroptosis by affecting the expression of ferroptosis-related proteins ACSL4, GPX4, and FTH1, thereby disrupting intracellular iron homeostasis and the GSH/GPX4 antioxidant defense system, ultimately leading to the accumulation of lipid peroxidation (LPO). Animal model demonstrated that B23 exhibited potent tumor suppression in an MDA-MB-231 xenograft model, achieving a tumor inhibition rate of 78.46 % at a dose of 4 mg/kg, without observable toxic side effects. In conclusion, B23 represents a promising ferroptosis inducer for the treatment of TNBC and warrants further investigation.

Bevacizumab, a monoclonal antibody targeting vascular endothelial growth factor A (VEGFA), is a widely used anti-angiogenic drug for malignancies. Its complex pharmacokinetics causes significant inter-individual concentration variations, making therapeutic drug monitoring (TDM) essential. Conventional liquid chromatography-tandem mass spectrometry (LC-MS/MS) and enzyme-linked immunosorbent assay (ELISA) for the TDM of bevacizumab suffer from limitations such as complex sample preparation and large sample volume requirements, highlighting the need for novel detection technologies. In this study, a surface plasmon resonance (SPR) biosensor for quantifying bevacizumab in human serum was developed. VEGFA was immobilized on a CM5 sensor chip, demonstrating high activity and specificity for bevacizumab. To mitigate nonspecific interference from the serum matrix, bovine serum albumin (BSA) was immobilized on the reference flow cell (Fc), serum dilution was optimized, and an appropriate sample diluent was selected. The sensor exhibited a detection range of 25-3200 ng mL-1. Intra-day and inter-day precision showed a coefficient of variation (CV) below 15% and an accuracy ranging between 85% and 115%. Finally, the biosensor was successfully applied to 15 clinical serum samples, showing significant correlation with the ELISA results. Compared to traditional methods, the SPR biosensor offers simpler preparation, faster analysis, and smaller sample volumes, which provides a new option for the TDM of bevacizumab.

Amikacin (AMK) is a semisynthetic antibiotic used in the treatment of gram-negative bacterial infections and has a narrow therapeutic index. Individualized treatment of amikacin under the guidance of therapeutic drug monitoring (TDM) is important to reduce the occurrence of toxicity and improve clinical efficacy. Current TDM techniques for AMK, such as chromatographic technology and immunoassay, are conducted in central labs using specialized equipment. However, the extensive time and costs involved hinder the application of AMK detection in medical practices. In the present study, a novel surface plasmon resonance (SPR) biosensor for detecting the concentration of AMK in human serum samples was developed and validated. The detection range of SPR method was 0.125 similar to 8 ng/ml, and the limit of detection was 0.035 +/- 0.004 ng/ml. The intra- and inter-day accuracy of SPR was 98 %-105 % and 97 %-110 %, respectively, which met the analytical requirements. The consistency evaluation with the high-performance liquid chromatography (HPLC) method shows that the SPR biosensor is reliable for quantification of AMK. This work is a foundation towards the development of a label-free, real-time, accurate, low cost, and simple method for future TDM, and will help clinicians to optimize dosing regimens thus to maximize the clinical effect and minimize the toxicity of these drugs.

Sepsis is a life-threatening systemic inflammatory response syndrome with high morbidity and mortality. A frequent complication is sepsis-associated encephalopathy (SAE), characterized by neurological impairments. Microglia pyrin domain-containing receptor 3 (NLRP3) inflammasome activation is a pivotal pathogenic mechanism in SAE. Microtubule-affinity regulating kinase 4 (MARK4) has been implicated in neuroinflammatory diseases, but its role in SAE and microglial NLRP3 inflammasome activation is unclear. Sophoridine (SRI), a MARK4 antagonist with anti-inflammatory and neuroprotective properties, has not yet been thoroughly investigated in SAE. Here, we used a cecum ligation puncture (CLP)-induced SAE mouse model and LPS + ATP or LPS + nigericin (Nig)-treated microglia, to assess the role of MARK4 in SAE. Molecular docking, Nissl staining, the enzyme-linked immunosorbent assay (ELISA), immunofluorescence, and behavioral tests were applied to assess SRI's effects on SAE. Our findings demonstrate that MARK4 is significantly upregulated in SAE. Knockdown of MARK4 effectively suppressed microglial pyroptosis and pro-inflammatory cytokine release by inhibiting NLRP3 inflammasome activation. SRI binds to MARK4 and enhances its stability. By suppressing MARK4 protein expression, SRI reduces NLRP3 inflammasome-mediated microglia inflammation. In vivo, SRI treatment increased survival, improved neurological behavioral deficits, attenuated blood-brain barrier disruption, and provided neuroprotective effects against brain injury. In summary, MARK4 contributes to microglial pyroptosis and neuroinflammation through NLRP3 pathway in SAE, resulting in neuronal injury and cognitive impairment. SRI alleviates SAE by targeting MARK4. These results suggest MARK4 as a potential therapeutic target and SRI as a promising candidate for treating SAE.

Small molecules that can bind to specific cells have broad application in cancer diagnosis and treatment. Screening large chemical libraries against live cells is an effective strategy for discovering cell-targeting ligands. The DNA-encoded chemical library (DEL or DECL) technology has emerged as a robust tool in drug discovery and has been successfully utilized in identifying ligands for biological targets. However, nearly all DEL selections have predefined targets, while target-agnostic DEL selections interrogating the entire cell surface remain underexplored. Herein, we systematically optimized a cell-based DEL selection method against cancer cells without predefined targets. A 104.96-million-member DEL was selected against MDA-MB-231 and MCF-7 breast cancer cells, representing high and low metastatic properties, respectively, which led to the identification of cell-specific small molecules. We further demonstrated cell-targeting applications of these ligands in cancer photodynamic therapy and targeted drug delivery. Finally, leveraging the DNA tag of DEL compounds, we identified α-enolase (ENO1) as the cell surface receptor of one of the ligands targeting the more aggressive MDA-MB-231 cells. Overall, this work offers an efficient approach for discovering cell-targeting small molecule ligands by using DELs and demonstrates that DELs can be a useful tool to identify specific surface receptors on cancer cells.

D-dimer is a protein fragment generated during the fibrin breakdown by plasmin, and it serves as a mature biomarker for diagnosing thrombotic disorders. A novel immunoassay method based on surface plasmon resonance (SPR) has been developed, validated, and successfully applied for the quantification of D-dimer in human plasma with high sensitivity and rapidity. In this methodological study, we investigated the activity and stability of the SPR biosensor, sample pre-processing, washing conditions, intra-day and inter-day precision and accuracy and detection parameters, including a limit of detection of 8.3 ng/mL, a detection range spanning from 31.25 to 4000 ng/mL, and a detection time of 20 min. We compared D-dimer plasma concentration determination results using SPR with a classical latex-enhanced immunoturbidimetric immunoassay in 29 healthy individuals and thrombotic patients, and both methods exhibited consistency. Furthermore, we propose a hypothesis about the relationship between the concentration of D-dimer and its molecular weight. With an increase in the D-dimer concentration in plasma, the D-dimer approaches its simplest form (190 kDa).

The rapid and accurate detection of illegal adulteration of chemical drugs into dietary supplements is a big challenge in the food chemistry field. Detection of compounds without a standard reference is even more difficult; however, this is a common situation. Here in this study, a novel "standard-free detection of adulteration" (SFDA) method was proposed and phosphodiesterase-5 inhibitor derivatives were used as an example to figure out the possibility and reliability of this SFDA method. After analysis by quadrupole coupled time of flight-tandem mass spectrometry detection and multivariable statistics, six common fragment ions were chosen to indicate whether adulteration was present or not, while 20 characteristic fragment ions indicated whether adulteration was by nitrogen-containing heterocycles or by anilines. Furthermore, the quantitative methods were conducted by high-performance liquid chromatography-tandem mass spectrometry. In a word, this strategy allows for a quick determination of dietary supplement adulteration without any need for standard materials, improving the efficacy of food safety testing.

Traditional Chinese Medicine (TCM) is a supremely valuable resource for the development of drug discovery. Few methods are capable of hunting for potential molecule ligands from TCM towards more than one single protein target. In this study, a novel dual-target surface plasmon resonance (SPR) biosensor was developed to perform targeted compound screening of two key proteins involved in the cellular invasion process of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2): the spike (S) protein receptor binding domain (RBD) and the angiotensin-converting enzyme 2 (ACE2). The screening and identification of active compounds from six Chinese herbs were conducted taking into consideration the multi-component and multi-target nature of Traditional Chinese Medicine (TCM). Puerarin from Radix Puerariae Lobatae was discovered to exhibit specific binding affinity to both S protein RBD and ACE2. The results highlight the efficiency of the dual-target SPR system in drug screening and provide a novel approach for exploring the targeted mechanisms of active components from Chinese herbs for disease treatment.