A Plasmonic Fluor-Lightened Microneedle Array Enables Ultrasensitive Multitarget Whole Blood Diagnosis of Anemia in A Paper Origami-Based Device.

This work reports a portable, origami-type paper device with a plasmonic fluor-labeled microneedle sensing module for the multiplexed quantification of anemia biomarkers in whole blood. Sequential steps, including serum separation, target enrichment, and multiplexed readout by a gel imager, are rapidly accomplished with the flexible and highly integrated device. The microneedle array enabled efficient sampling of trace targets from ng mL-1 to pg mL-1 level. Combined with the plasmonic fluor label, the signal is improved by ≈7.6 folds compared with the flat substrate-based assay. The device is applied to simultaneously quantify hemoglobin (Hb), ferritin, folic acid (FA), and vitamin B12 (VB12 ), which are four anemia biomarkers distributed in different environments with different concentration ranges. Featured by the small sample volume (150 µL), short assay time (20 min), low cost (2 $), robust stability, and user-friendliness, the device is promising for the rapid and accurate diagnosis of anemia in real practice.

Ultrasonography characteristics of cystic components in primary salivary gland tumors.

OBJECTIVES: The present study aimed to characterize the ultrasonography (US) features of cystic components in salivary gland tumors (SGTs). MATERIALS AND METHODS: A total of 207 patients (218 lesions) with pathologically confirmed primary SGTs were analyzed. Preoperative US revealed the presence of cystic components in lesions. Lesion size, shape, margin, and US findings of the cystic components, including number, distribution, margin, occupying rate, and internal characteristics, were evaluated. RESULTS: Similarities were observed between the US performance of benign SGTs (B-SGTs) and malignant SGTs (M-SGTs) with cystic components. Differences in sex and age of patients, number, distribution, and internal characteristics of cystic components were statistically significant. For SGTs with cystic components, the proportions of M-SGTs to ill-defined margins (P = 0.002), eccentric distribution (P = 0.019), and none of the internal characteristics (P = 0.019) were significantly higher than those of B-SGTs. Younger age (P = 0.001), eccentric distribution (P = 0.034) and ill-defined margin (P < 0.001) were risk factors for diagnosing M-SGTs. Cystic component features needed to be combined with lesion indicators (border and shape) to improve diagnostic sensitivity. CONCLUSIONS: US features of the B-SGTs and M-SGTs were significantly different. Cystic component is of interest in the US-related differential diagnosis of B-SGT and M-SGT. CLINICAL RELEVANCE: Cystic components are potentially valuable in the differential diagnosis of B-SGTs and M-SGTs on US.

Transcriptional analysis of Bemisia tabaci MEAM1 cryptic species under the selection pressure of neonicotinoids imidacloprid, acetamiprid and thiamethoxam.

BACKGROUND: Neonicotinoids are widely applied in the control of the destructive agricultural pest Bemisia tabaci, and resistance against these chemicals has become a common, severe problem in the control of whiteflies. To investigate the molecular mechanism underlying resistance against nenonicotinoids in whiteflies, RNA-seq technology was applied, and the variation in the transcriptomic profiles of susceptible whiteflies and whiteflies selected by imidacloprid, acetamiprid and thiamethoxam treatment was characterized. RESULTS: A total of 90.86 GB of clean sequence data were obtained from the 4 transcriptomes. Among the 16,069 assembled genes, 584, 110 and 147 genes were upregulated in the imidacloprid-selected strain (IMI), acetamiprid-selected strain (ACE), and thiamethoxam (THI)-selected strain, respectively, relative to the susceptible strain. Detoxification-related genes including P450s, cuticle protein genes, GSTs, UGTs and molecular chaperone HSP70s were overexpressed in the selected resistant strains, especially in the IMI strain. Five genes were downregulated in all three selected resistant strains, including 2 UDP-glucuronosyltransferase 2B18-like genes (LOC 109030370 and LOC 109032577). CONCLUSIONS: Ten generations of selection with the three neonicotinoids induced different resistance levels and gene expression profiles, mainly involving cuticle protein and P450 genes, in the three selected resistant whitefly strains. The results provide a reference for research on resistance and cross-resistance against neonicotinoids in B. tabaci.

Imaging identification of primary mammary analogue secretory carcinoma and acinic cell carcinoma in major salivary glands.

OBJECTIVE: The present study aimed to characterize and differentiate the ultrasonography (US) and computed tomography (CT) features of mammary analogue secretory carcinoma (MASC) and acinic cell carcinoma (AciCC). METHODS AND PATIENTS: A total of 83 patients with clinically proven MASC and AciCC were analyzed. The following characteristics were assessed on US, CT, and magnetic resonance imaging: lesion size, shape, margin, echogenicity, echotexture, cystic components, posterior echo, vascularity, density, degree of enhancement, enhancement pattern, signal intensity (SI) on T1- and T2-weighted images (WI), hemorrhages, and lymph node enlargement. RESULTS: Similarities were observed between the imaging performance of MASC and AciCC. Differences between the two characteristics of shape on US and cystic components on CT were statistically significant. The proportion of MASC to regular shape on US (p = 0.006) and cystic components on CT (p = 0.027) was significantly higher than that of AciCC. Regular shape on US had the highest sensitivity in the identification of MASC and AciCC, while regular shape on US + cystic component on CT had the highest specificity. CONCLUSIONS: The shape on US and cystic components on CT are key characteristics for distinguishing MASC and AciCC.

A visual and reversible nanoprobe for rapid and on-site determination of hexavalent chromium and lysine based on dual-emission carbon quantum dots coupled with smartphone.

A straightforward, largely instrument-free, smartphone-based analytical strategy for hexavalent chromium and lysine (Lys) on-site detection via exploitation of dual-emission carbon quantum dots (DECQDs) has been demonstrated. DECQDs show dual-emission peaks at 439 and 630 nm with the excitation at 375 nm. As a dual-mode detection probe, the fluorescence and ultraviolet adsorption spectra of DECQDs vary with hexavalent chromium concentrations. Most importantly, Lys can restore the fluorescence of the hexavalent chromium added DECQD nanoprobe and change the color of the probe under natural light. At the same time, based on the participation of smartphones, the prepared DECQD probes favor the establishment of visual smart sensors that can also be used for the in-situ detection of targets. The on-site quantitative analysis exhibited a linear range of 5.3-320 µM with a detection limit of 1.6 µM towards Cr(VI) and the differentiation of Lys variation from 1 to 75 mM with a detection limit of 0.3 mM. The probe has been applied for the first time to enable vision-based colorimetric in complex samples such as water, milk and egg. The recoveries of Cr(VI) and Lys in real samples were between 90 and 104%, and the relative standard deviation (RSD) was as low as 0.4%. This work offers new perspectives for fundamental understanding and new design of functional luminescent materials that are applicable for food-safety and rapid and intelligent inspection. A straightforward, large instrument-free, smartphone-based analytical strategy with dual-emission carbon quantum dots was developed for hexavalent chromium and Lys on-site detection via fluorescent and colorimetric twofold readout measure.

Identification and Characterization of Specific Nanobodies against Trop-2 for Tumor Targeting.

Trophoblast cell-surface antigen 2 (Trop-2) is a tumor-associated antigen that is connected with the development of various tumors and has been identified as a promising target for tumor immunotherapy. To date, the immunotherapy against Trop-2 mainly relies on the specific targeting by monoclonal antibody in antibody-drug conjugate (ADC). Alternatively, the single domain antibodies of nanobodies (Nbs) possesses unique properties such as smaller size, better tissue penetration, etc., to make them good candidates for tumor targeting. Thus, it was proposed to develop anti-Trop-2 Nbs for tumor targeting in this study. Generally, three consecutive rounds of bio-panning were performed against immobilized recombinant Trop-2, and yielded three Nbs (Nb60, Nb65, and Nb108). The affinity of selected Nbs was determined in the nanomolar range, especially the good properties of Nb60 were verified as a promising candidate for tumor labeling. The binding to native Trop-2 was confirmed by flow cytometry against tumor cells. The inhibitory effects of the selected Nbs on tumor cell proliferation and migration were confirmed by wound healing and Transwell assay. The clear localization of the selected Nbs on the surface of tumor cells verified the potent labeling efficiency. In conclusion, this study provided several Nbs with the potential to be developed as targeting moiety of drug conjugates.

Integration of Metal-Organic Frameworks with Bi-Nanoprobes as Dual-Emissive Ratiometric Sensors for Fast and Highly Sensitive Determination of Food Hazards.

Functional nanoprobes which detect specific food hazards quickly and simply are still in high demand in the field of food-safety inspection research. In the present work, a dual-emission metal-organic framework-based ratiometric fluorescence probe was integrated to detect Cu2+ and Pb2+ with rapidness and ease. Specifically, quantum dots (QDs) and carbon quantum dots (CQDs) were successfully embedded into zeolitic imidazolate framework-67 (ZIF-67) to function as a novel ratiometric fluorescent sensing composite. The ratiometric fluorescence signal of CQDs/QDs@ZIF-67 was significantly aligned with the concentration of metal ions to give an extremely low detection limit of 0.3324 nM. The highly sensitive and selective CQDs/QDs@ZIF-67 composite showed potential for the rapid and cost-effective detection of two metal ions.

A Universal Biofilm Reactor Sensor for the Determination of Biochemical Oxygen Demand of Different Water Areas.

In this study, we developed a simple strategy to prepare a biofilm reactor (BFR) sensor for the universal biochemical oxygen demand (BOD) determination. The microorganisms in fresh water were domesticated by artificial seawater with different salinity gradients successively to prepare the BFR sensor. The prepared BFR sensor exhibits an efficient ability to degrade a variety of organic substances. The linear range of BOD determination by the BFR sensor is 1.0-10.0 mg/L-1 with a correlation coefficient of 0.9951. The detection limit is 0.30 mg/L according to three times of signal-to-noise ratio. What is more, the BFR sensor displayed excellent performances for the BOD determination of different water samples, including both fresh water and seawater. The 16S-rRNA gene sequencing technology was used to analyze the microbial species before and after the domestication. The results show that it is a general approach for the rapid BOD determination in different water samples.

PEITC triggers multiple forms of cell death by GSH-iron-ROS regulation in K7M2 murine osteosarcoma cells.

Phenethyl isothiocyanate (PEITC) is an isothiocyanate that largely exists in cruciferous vegetables and exhibits chemopreventive and chemotherapeutic potential against various cancers. However, it is little known about the molecular mechanisms of its antitumor action against osteosarcoma, which is the second highest cause of cancer-related death in children and adolescents. In this study, we investigated the effects of PEITC on K7M2 murine osteosarcoma both in vitro and in vivo. We found that treatment with PEITC dose-dependently inhibited the viability of K7M2 murine osteosarcoma cells with an IC50 value of 33.49 µM at 24 h. PEITC (1, 15, 30 µM) dose-dependently inhibited the cell proliferation, caused G2/M cell cycle arrest, depleted glutathione (GSH), generated reactive oxygen species (ROS), altered iron metabolism, and triggered multiple forms of cell death, namely ferroptosis, apoptosis, and autophagy in K7M2 cells. We further revealed that PEITC treatment activated MAPK signaling pathway, and ROS generation was a major cause of PEITC-induced cell death. In a syngeneic orthotopic osteosarcoma mouse model, administration of PEITC (30, 60 mg/kg every day, ig, for 24 days) significantly inhibited the tumor growth, but higher dose of PEITC (90 mg/kg every day) compromised its anti-osteosarcoma effect. Histological examination showed that multiple cell death processes were initiated, iron metabolism was altered and MAPK signaling pathway was activated in the tumor tissues. In conclusion, we demonstrate that PEITC induces ferroptosis, autophagy, and apoptosis in K7M2 osteosarcoma cells by activating the ROS-related MAPK signaling pathway. PEITC has promising anti-osteosarcoma activity. This study sheds light on the redox signaling-based chemotherapeutics for cancers.

Simulation and observation for volume emission rates emitted from O2(0-1) and O(1S) nightglow in northwest China.

Being susceptible to the change of atmospheric conditions, the volume emission rate (VER) is very suitable to be used as a light source by passive remote sensing for measuring atmospheric wind and temperature. Thus, the VERs emitted from O2(0-1) and O(S1) of the nightglow at 80-120 km are studied in this paper. Based on the Naval Research Laboratory Mass Spectrometer Incoherent Scatter (NRLMSISE-00) model data and the ground-based airglow imaging interferometer (GBAII) instrument observation for a local time and place, simulated VER profiles represented by four layers are obtained for the nightglow of O2(0-1) and O(S1). The O2(0-1) nightglow model peak values at 94 km on 6 December 2013 and 8 November 2011 are 8111 photons·cm-3·s-1 and 8406 photons·cm-3·s-1, respectively; however, the O(S1) VER peak at a higher altitude of about 96 km on 18 December 2011 is only 338 photons·cm-3·s-1. The upper atmospheric VER values have been derived to transfer into the ground-based detected column intensities by our GBAII prototype. The calculated column integrated emission rates (IERs) of O2(0-1) for 0° and 45° zenith angles are 1.48×107 and 1.91×107 photons·cm-2·s-1, respectively; the calculated column IERs of O(S1) are 5.53×105 and 7.03×105 photons·cm-2·s-1, respectively. Correspondingly, the detected column IERs obtained by GBAII are 2.43×107 for O2(0-1) and 6.57×105 photons·cm-2·s-1 for O(S1).

Immunomodulation of nutritional formula containing epigallocatechin-3-gallate, ginseng extract, and polydextrose on inflammation and macrophage polarization.

Single nutrient likes polyphenol or dietary fiber have been exhaustively investigated to validate their positive intervention in health or disease. Meanwhile, the common interaction of inner systems with the nutrient complex has not been well elucidated, which raises the scientific issue of the modulatory effect of the nutrient complex on immunity. The representative prebiotics of epigallocatechin-3-gallate (EGCG), ginseng extract, and polydextrose (PDX) were selected on behalf of the classification of polyphenol, flavone or polysaccharides, and dietary fiber to generally cover the daily food intake in this study to explore their intervention in inflammation and macrophage polarization. The intervention of selected nutrients on inflammation and macrophage polarization has been evaluated against macrophages to unveil their comprehensive effects. The synergistic effect of selected nutrients was demonstrated by inhibiting M1 macrophage polarization and the promotion of M2 macrophage polarization. Then, the nutrient formula was set up to verify the intervention effect, and the results revealed the significant inhibition of cell inflammation and the effect on cell proliferation through promoting the cell cycle in the G2 phase. The nutrient complex could inhibit M1 macrophage polarization to inhibit M1-mediated inflammation and promote M2 macrophages for anti-inflammatory effect and enhance cell phagocytosis. Moreover, the varied intervention effects of the nutrient complex with different formulas could be summarized. In general, the formula containing EGCG, ginseng extract, and PDX was demonstrated to possess an enhanced immunomodulatory effect on cell inflammation and macrophage polarization, which could potentially inspire the investigation of complex nutrients in health and diseases.

Nanobody-Based Electrochemical Immunoassay for Sensitive Detection of Peanut Allergen Ara h 1.

Peanut is widely used for food supplementation with potential allergic reactions in infants and adults, which prompted the development of reliable and accurate detection of peanut allergens with emphasis on Ara h 1. In this study, a nanobody (Nb)-based micro-total electrochemical immunoassay (Nb-µTEI) was proposed to be generated. Generally, an alpaca was immunized with Ara h 1 to yield a Nb reservoir for selection of four specific Nbs. Nb-mediated immunocapturing allowed the identification of the target as Ara h 1. The Nb-based electrochemical immunoassay was developed by constructing a capturing electrode with cycles of signal enhancement. After construction of the capturing electrode, Nb152 with HA-tag was directly applied to connect immobilized anti-HA IgG for the capture of different concentrations of Ara h 1, which was labeled by biotinylated Nb152 to facilitate signal development with alkaline phosphatase conjugated streptavidin (SA-ALP). A linear range from 4.5 to 55 ng/mL was acquired with LOD and LOQ of 0.86 and 2.10 ng/mL, respectively, with an 11-fold increase of the sensitivity compared with the established sandwich ELISA. The dedicated immunoassay was verified by detecting the antigen spiked in food samples and demonstrated the successful conjugation of Nb with advanced detecting techniques.

Hollow-structured molecularly imprinted polymers enabled specific enrichment and highly sensitive determination of aflatoxin B1 and sterigmatocystin against complex sample matrix.

The biotoxins with high toxicity have the potential to be manufactured into biochemical weapons, seriously threatening international public security. Developing robust and applicable sample pretreatment platforms and reliable quantification methods has been recognized as the most promising and practical approach to solving these problems. Through the integration of the hollow-structured microporous organic networks (HMONs) as the imprinting carriers, we proposed a molecular imprinting platform (HMON@MIP) with enhanced adsorption performance in terms of specificity, imprinting cavity density as well as adsorption capacity. The HMONs core of MIPs provided a hydrophobic surface that enhanced the adsorption of biotoxin template molecules during the imprinting process, resulting in an increased imprinting cavity density. The HMON@MIP adsorption platform could produce a series of MIP adsorbents by changing the biotoxin template, such as aflatoxin and sterigmatocystin, and showed promising generalizability. The limits of detection (LOD) of the HMON@MIP-based preconcentration method for AFT B1 and ST were 4.4 and 6.7 ng L-1, respectively, and the method was applicable to food sample with satisfied recoveries of 81.2-95.1%. And the specific recognition and adsorption sites left on HMON@MIP by the imprinting process can achieve outstanding selectivity for AFT B1 and ST. The developed imprinting platforms hold great potential for application in the identification and determination of various food hazards in complex food sample matrices and contribute to precise food safety inspection.

The Effect of Mother Phubbing on Young Children's Emotional and Behavioral Problems: A Moderated Mediation Model of Mother-Child Attachment and Parenting Stress.

Phubbing-the act of ignoring someone physically present in favor of a mobile phone-is increasingly prevalent in families, and mothers' phubbing behaviors may have a particularly important effect on young children's development. Accordingly, this study explores the mediating role of mother-child attachment in the relationship between mother phubbing and children's emotional and behavioral problems, as well as the role of maternal parenting stress in moderating the mediation effect. A total of 988 mothers of young children (mean age = 4.93, SD = 0.94) were surveyed using four scales, and the resulting data was statistically analyzed. The study found that (1) mother phubbing was significantly and positively correlated with children's emotional and behavioral problems (r = 0.19, p < 0.01), (2) mother-child attachment mediated the relationship between mother phubbing and children's emotional and behavioral problems, and (3) the relationship between mother-child attachment and children's emotional and behavioral problems was moderated by maternal parenting stress. The present study offers fresh evidence of how mother phubbing affects young children's emotional and behavioral difficulties. The need to reduce maternal parental stress and buffer mothers from its effects are highlighted as vital factors in promoting secure mother-child attachment and alleviating young children's problems.

Hollow-Structured Microporous Organic Networks Adsorbents Enabled Specific and Sensitive Identification and Determination of Aflatoxins.

Aflatoxin (AFT) contamination, commonly in foods and grains with extremely low content while high toxicity, has caused serious economic and health problems worldwide. Now researchers are making an effort to develop nanomaterials with remarkable adsorption capacity for the identification, determination and regulation of AFT. Herein, we constructed a novel hollow-structured microporous organic networks (HMONs) material. On the basis of Fe3O4@MOF@MON, hydrofluoric acid (HF) was introduced to remove the transferable metal organic framework (MOF) to give hollow MON structures. Compared to the original Fe3O4@MOF@MON, HMON showed improved surface area and typical hollow cavities, thus increasing the adsorption capacity. More importantly, AFT is a hydrophobic substance, and our constructed HMON had a higher water contact angle, greatly enhancing the adsorption affinity. From that, the solid phase extraction (SPE-HPLC) method developed based on HMONs was applied to analyze four kinds of actual samples, with satisfied recoveries of 85-98%. This work provided a specific and sensitive method for the identification and determination of AFT in the food matrix and demonstrated the great potential of HMONs in the field of the identification and control of mycotoxins.

Instantaneous discharge characteristics and its methane ignition mechanism of coal mine rock damage.

The goaf is the main site of gas explosions in coal mines. A large number of accidents indicate that the occurrence of gas explosion in the goaf is related to the deformation and fracture of the goaf roof. An experimental system was constructed to study the instantaneous discharge characteristics and its ignition mechanism caused by rock damage. The research results reveal that different rocks produce avalanche-like dust clouds ejected outward during the fracture process, and the generation process takes an extremely short time. This dust cloud is not caused by rock fragments heated to incandescence because of friction. The electric sparks produced in the process of rock rupture and the appearance of instantaneous discharge are related not only to the quartz content of the rock, but also to the compressive strength of the rock. The piezoelectric effect of the rock becomes the key factor for the strength or the generation of an electric spark. The instantaneous discharge leaded to the collision of high-energy electrons released during the fracture process with air molecules, ionising, and breaking down the air. When the surrounding medium is gas air within the explosion limit, ionisation is formed and causes gas explosions.

Preparation and Band Gap Characteristics of Composite Film/Substrate Instability System.

Soft materials such as biological tissues are prone to deformation and generate different stable structures under external stimulation. This property is widely used to create tunable patterns, and the tuning of the wrinkling patterns can be applied to the control of elastic waves. In this paper, the wrinkling modes of film/substrate systems with different geometric dimensions and material parameters were studied. It is verified by numerical simulation that the elastic wave band gaps corresponding to the two wrinkling modes can be effectively superposed in one system, and the experimental samples with two wrinkling modes coexisting in one system were prepared by parameter optimization and a moisture-curing process. A vibration test showed that the hybrid system could effectively suppress the propagation of elastic waves. Combined with engineering needs, the wrinkling system under different loading conditions was studied, which provides a design guide for widening and regulating the elastic wave band gap.

Protective effects of bioactive peptides in foxtail millet protein hydrolysates against experimental colitis in mice.

It is of great significance to develop a dietary intervention strategy to prevent inflammatory bowel disease (IBD). A millet-rich diet can ameliorate IBD, but the active ingredients and mechanisms remain to be studied. Our results showed that the oral administration of foxtail millet protein hydrolysates (FMPH) reduced the disease activity index (DAI) score and improved the colon symptoms of dextran sulfate sodium (DSS)-induced colitis mice. FMPH reduced the serum LPS level, increased intestinal ZO-1 and occludin expression, inhibited NF-κB phosphorylation, and reduced the levels of TNF-α and IL-6. Further, FMPH inhibited Th17 cell differentiation, and inhibited inflammasome activation and IL-1ß expression through the NLRP3/ASC/caspase-1 pathway. The results on Caco-2 cells confirmed the role of FMPH on tight junction and inflammasomes activation. A total of 2620 peptides were identified in FMPH by UPLC-MS/MS, of which 22 peptides were predicted as potential biopeptides, and the key sequences were LPF, ANP, PY, YW, and IPP. This study supports the effect of a diet rich in millet on the improvement of IBD and provides a scientific basis for the use of millet protein as a functional food to improve intestinal inflammation.

Sweeteners Maintain Epithelial Barrier Function Through the miR-15b/RECK/MMP-9 Axis, Remodel Microbial Homeostasis, and Attenuate Dextran Sodium Sulfate-Induced Colitis in Mice.

Non-nutritive sweeteners are the most widely used food additives designed to provide sweetness and reduce caloric intake. Studies have confirmed a link between sweeteners and colitis, yet supporting scientific data remain exiguous and controversial. In this study, three common sweeteners (Saccharin sodium, Stevioside, and Sucralose) in acceptable daily intake dosage were added to water in order to determine their effects on dextran sodium sulfate-induced colitis in mice. Our results show that the three sweeteners meliorate colitis to varying degrees─Saccharin exerts the most pronounced effect, followed by Stevioside and Sucralose. Intake of sweeteners alleviates colitis symptoms, alters gut microbiota, reshapes the TH17/Treg balance, protects the intestinal barrier, and reduces inflammation. Most significantly, sweeteners can enhance the abundance of Mucispirillum and Alistipes, which are conducive to colitis recovery, and upregulate the expression of E-cadherin through the miR-15b/RECK/MMP-9 axis to improve intestinal barrier integrity. Moreover, by inhibiting the MMP-9/AKT/NF-κB pathway, inflammation is relieved, as reflected in the restoration of the Th17/Treg balance. Our results link the consumption of sweeteners to the remission of colitis, which provides new scientific evidence for the safe use of sweeteners.

Exploration of Specific Nanobodies As Immunological Reagents to Detect Milk Allergen of ß-Lactoglobulin without Interference of Hydrolytic Peptides.

Milk proteins are widely used for food supplementation, despite the potential risk of food allergy, especially against ß-lactoglobulin (BLG), which makes BLG surveillance critical. Possible interaction of detecting antibodies with BLG-derived peptides will result in unprecise inspection. Thus, in this study, it was proposed to generate nanobodies (Nbs) and validate the immunological detection of intact BLG rather than hydrolytic peptides. Nbs were successfully retrieved and characterized with high stability and target specificity. A competitive enzyme-linked immunosorbent assay (cELISA) was developed with a linear range from 39 to 10,000 ng/mL and a detection limit (LOD) of 4.55 ng/mL, with a recovery of 86.30%-95.09% revealed by analysis of spiked samples. Meanwhile, a sandwich ELISA (sELISA) was established with Nb82 and BLG polyclonal antibody (pAb-BLG) providing a linear range from 29.7 to 1250 ng/mL and an LOD of 13.82 ng/mL with a recovery of 87.82%-103.97%. The interaction of selected Nbs with BLG-derived peptides was investigated by Nb structure modeling and BLG docking. No binding on hydrolytic peptides was revealed, confirming the precision of Nb-mediated immunoassays. In summary, this study successfully identified BLG-specific Nbs for immunoassay development and guaranteed the monitoring of intact BLG without interference of hydrolytic peptides, providing experimental evidence that our Nbs recognize intact food allergen.