Small Molecule Pollutants Research Articles

Fabrication of Raman reporter molecule–embedded magnetic SERS tag for ultrasensitive immunochromatographic monitoring of Cd ions and clenbuterol in complex samples

A rapid, simple but sensitive analytical method for the simultaneous monitoring of heavy metal ions and illegal additives in complex samples remains lacking. We reported a difunctional capture–detection surface-enhanced Raman scattering (SERS) immunochromatographic assay (ICA) for the direct detection of the heavy metal residue Cd ion (Cd2+) and illegal additive clenbuterol (CLE) with high sensitivity and specificity herein. A novel magnetic SERS tag (Fe@RAu) with a Raman reporter molecule–embedded raspberry-like Au shell was fabricated via a one-step reduction reaction that not only simplified the fabrication of common magnetic SERS nanomaterials but also improved SERS activity to increase detection sensitivity. The difunctional SERS–ICA enabled the fast magnetic enrichment of the target small molecules in complex specimens and ultrasensitive and quantitative determination of CLE/Cd2+ on the two test lines of the ICA system. The limits of detection of the developed method for CLE and Cd2+ were as low as 0.48 and 1.88 pg/mL, respectively, and about 2000 and 500 times lower than those of conventional AuNP-based ICA strips. The excellent practicability of Fe@RAu–ICA for real food samples analysis was also demonstrated, revealing the potential of the proposed assay for rapid and on-site screening of small-molecule pollutants.

Analysis of okadaic acid using electrochemiluminescence imaging on microfluidic biosensing chip

The sensitivity and specificity of electrochemiluminescence (ECL)-based biosensor directly rely on the property of luminophor, the type of sensing carriers and the effectiveness of signal amplification used in the sensor design, which poses a major challenge to manage these elements simultaneously. In this work, an aggregation-induced electrochemiluminescence (AIECL) microfluidic sensing chip using 4′,4″,4‴,4‴'-(ethene-1,1,2,2-tetrayl)tetrabiphenyl-4-carboxylic acid (TPE)-derived hafnium-based metal-organic framework (Hf-MOF) as emitter was developed. An easily overlooked marine pollutant, okadaic acid (OA) with different concentrations ranging from 5.00 ng/mL to 1.50 × 104 ng/mL at the electrode is visualized imaging benefit from high luminescence efficiency of Hf-MOF coupled the rolling circle amplification strategy assisted by trans-cleavage activity of CRISPR/Cas12a. These highlights will solve the long-lasting task in the accurate analysis of small molecule pollutants, which can be able to provide more worthy reference solution about construction of novel ECL luminophor and signal extraction of low-abundance disease-related biomarkers.

Core–satellite-structured magnetic nanozyme enables the ultrasensitive colorimetric detection of multiple drug residues on lateral flow immunoassay

BackgroundExcessive use of veterinary drugs causes severely environmental pollution and agricultural pollution, and poses great threat to human health. A simple method for the rapid, highly sensitive, and on-site monitoring of veterinary drug residues in complex samples remains lacking. ResultsIn this study, we propose a catalytically enhanced colorimetric lateral flow immunoassay (LFA) based on a novel core–satellite-structured magnetic nanozyme (Fe–Au@Pt) that can simultaneously and quantitatively detect three common veterinary drugs, namely, gentamicin (GM), streptomycin (STR), and clenbuterol (CLE), within a short testing time (<30 min). The Fe–Au@Pt nanozyme was simply prepared through the self-assembly of numerous Au@Pt nanoparticles on a large Fe3O4 core via electrostatic adhesion, which exhibited the advantages of high peroxidase-like activity, strong magnetic responsiveness, and multiple catalytic sites. Under the dual-signal amplification effect of magnetic enrichment and catalytic enhancement, the proposed nanozyme-LFA allowed the multiplex detection of STR, CLE, and GM with detection limits of 10.1, 6.3, and 1.1 pg/mL, respectively. SignificanceThe developed Fe–Au@Pt–LFA achieves direct, simultaneous, and accurate detection of three target drugs in food samples (honey, milk, and pork). The proposed assay shows great potential for application in the real-time monitoring of small-molecule pollutants in complex environment.

Sustainable B12-Dependent Dehalogenation of Organohalides in E. coli.

Microbial bioremediation can provide an environmentally friendly and scalable solution to treat contaminated soil and water. However, microbes have yet to optimize pathways for degrading persistent anthropogenic pollutants, in particular organohalides. In this work, we first expand our repertoire of enzymes useful for bioremediation. By screening a panel of cobalamin (B12)-dependent reductive dehalogenases, we identified previously unreported enzymes that dechlorinate perchloroethene and regioselectively deiodinate the thyroidal disruptor 2,4,6-triiodophenol. One deiodinase, encoded by the animal-associated anaerobe Clostridioides difficile, was demonstrated to dehalogenate the naturally occurring metabolites L-halotyrosines. In cells, several combinations of ferredoxin oxidoreductase and flavodoxin extract and transfer low-potential electrons from pyruvate to drive reductive dehalogenation without artificial reductants and mediators. This work provides new insights into a relatively understudied family of B12-dependent enzymes and sets the stage for engineering synthetic pathways for degrading unnatural small molecule pollutants.

Dosage-sensitive and simultaneous detection of multiple small-molecule pollutants in environmental water and agriproducts using portable SERS-based lateral flow immunosensor

The co-contamination of pesticide residues and mycotoxins in agricultural products is a global concern, with the potential for cumulative and synergistic damaging effects, imposing substantial health and economic burdens to the public. The dosage-sensitive and simultaneous detection of multiple pollutants, with a heightened sensitivity in real samples, poses a significant demand and challenge. Herein, we propose a portable detection method integrating surface-enhanced Raman scattering (SERS)-with lateral flow immunoassay (LFIA), offering high sensitivity and multiplex analysis capabilities. This approach enables the simultaneous detection of imidacloprid (IMI), pyraclostrobin (PYR) and aflatoxin B1 (AFB1) through a single test strip. Utilizing the immune-specific binding between antigen and antibodies, we immobilised antibody- conjugated SERS nanotags on three test lines of the strips to generate Raman signal amplification in the proposed biosensor. Accurate quantitative analysis was performed by measuring the SERS signal intensity on the test lines. The limits of detection were 8.6 pg/mL for IMI, 97.4 pg/mL for PYR and 8.9 pg/mL for AFB1, exhibiting sensitivities 12-fold, 102-fold and11-fold higher than the colorimetric signals, respectively. Importantly, the SERS-LFIA immunosensor demonstrated robust performance when applied to real samples, yielding recoveries ranging from 86.16 % to 115.0 %, with relative standard deviation values below 8.67 %. These results underscore the excellent stability, high selectivity and reliability the proposed SERS-LFIA immunosensor. Consequently, it holds promise for the detection of multiple pesticides and mycotoxins in both environmental and agricultural samples.

Dual-color MoS2@QD nanosheets mediated dual-mode lateral flow immunoassay for flexible and ultrasensitive detection of multiple drug residues

The sensitive on-site monitoring of multiple agricultural drugs in complex real media is critical to human health but remains a challenge. In this paper, we reported the design of film-like dual-mode nanolabel (MTQD), using molybdenum disulfide (MoS2) nanosheet-loaded numerous quantum dots (QDs) to generated co-enhanced colorimetric-fluorescence signals, and demonstrated its powerful capabilities in competitive lateral flow immunoassay (LFA) for flexible, ultrasensitive, and multiplex detection of four kinds of common agricultural drugs. Three layers of red and green QDs were continuously coated onto the surface of MoS2 to suppress the inner filter effect of two-dimensional nanomaterials effectively and greatly improve the specificity of the LFA system for multitarget analysis. The dual-mode LFA combined with a low-cost dual-fluorescence reading instrument can flexibly and simultaneously monitor two pesticides (clothianidin, carbendazim) and two veterinary drugs (kanamycin, chloramphenicol) in complex food and experimental samples with low detection limits of 0.1 ng/mL level in the observation of colorimetric signals and showed at least a 46-fold improvement in sensitivity in the sensing of fluorescence signals. This colorimetric dual-fluorescence platform greatly broadens the application range of existing LFA techniques without increasing testing time, thus showing great application prospects for the point-of-care testing of small-molecule pollutants in different situations.

Ultrasensitive Ratiometric Electrochemiluminescence Sensor with an Efficient Antifouling and Antibacterial Interface of PSBMA@SiO2-MXene for Oxytetracycline Trace Detection in the Marine Environment.

The sensitivity and accuracy of electrochemiluminescence (ECL) sensors for detecting small-molecule pollutants in environmental water are affected not only by nonspecific adsorption of proteins and other molecules but also by bacterial interference. Therefore, there is an urgent need to develop an ECL sensor with antifouling and antibacterial functions for water environment monitoring. Herein, a highly efficient antifouling sensing interface (PSBMA@SiO2-MXene) based on zwitterionic sulfobetaine methacrylate (SBMA) antifouling nanospheres (NPs) and two-dimensional MXene nanosheets was designed for the sensitive detection of oxytetracycline (OTC), an antibiotic small-molecule pollutant. Specifically, SBMA with good hydrophilicity and electrical neutrality was connected to SiO2 NPs, thus effectively reducing protein and bacterial adsorption and improving stability. Second, MXene with a high specific surface area was selected as the carrier to load more antifouling NPs, which greatly improves the antifouling performance. Meanwhile, the introduction of MXene also enhances the conductivity of the antifouling interface. In addition, a ratio-based sensing strategy was designed to further improve the detection accuracy and sensitivity of the sensor by utilizing Au@luminol as an internal standard factor. Based on antifouling and antibacterial interfaces, as well as internal standard and ratiometric sensing strategies, the detection range of the proposed sensor was 0.1 ng/mL to 100 μg/mL, with a detection limit of 0.023 ng/mL, achieving trace dynamic monitoring of antibiotics in complex aqueous media.

Forging Fenton-like system to functionalize electron/hole enrichment regions for ATZ photocatalytic degradation

Independent functional regions for electrons and holes enrichment are usually driven by the construction of multiple heterostructures, which is difficult to achieve fine-controlling, resulting in the limited photocatalytic activity. TiO2-based photocatalysts with suitable forbidden band widths are promising for wide application prospects, where the construction of Schottky heterojunctions is the trick to achieve directional migration of electrons and holes. Here, a regionalized surface modification method of H–TiO2 with oxygen vacancy (Ov) and defects for efficient carrier separation was reported, in which interspersed Ag nanoparticles and SiO2 layers serve as electron/hole enrichment regions, respectively. Meanwhile, SiO2 can be served as adsorption sites for photocatalytic oxidation of small-molecule pollutants, while Ag nanoparticles with H+ adsorption capacity and reasonable guide band position can be used as an engine of Fenton-like reactions to cyclically generate the active substance H2O2 to participate in the reduction reaction. The degradation efficiency of Ag–SiO2–H–TiO2 (ASHT) can be increased by 159% for pesticide atrazine (ATZ). The superoxide radicals generated by electrons and oxygen in ASHT also generate H2O2, which promotes the centralized generation of hydroxyl radicals to improve photocatalytic performance. The structural design strategy of ASHT provides a new concept for the process of ring-opening reactions of benzene-containing contaminants in water and conversion to small molecules which will eventually be rapidly mineralized.

Portable multichannel detection instrument based on time-resolved fluorescence immunochromatographic test strip for on-site detecting pesticide residues in vegetables

In this work, a portable multichannel detection instrument based on time-resolved fluorescence immunochromatographic test strip (TRFIS) was proposed for on-site detecting pesticide residues in vegetables. Its hardware consisted of a silicon photodiode and excitation light source array, a mainboard of the lower machine with STMicroelectronics 32 (STM32) and a linear stepping motor. While detecting, cardboard with 6-channel TRFIS was pulled into the cassette by the stepping motor. The peak area of the test (T) line and control (C) line of each TRFIS was sampled and calculated by software, then the concentration of the detected pesticide was obtained according to the ratio of the T to C value. This instrument could sample 6-channel TRFIS within 30 s simultaneously, and it exhibited excellent accuracy with a 2.5% average coefficient of variation for each channel (n = 12). In addition, the TRFIS was constructed by using europium oxide time-resolved fluorescent microspheres to label the monoclonal antibody against acetamiprid and form a fluorescent probe, which was fixed on the binding pad. The TRFIS was used for the detection of acetamiprid in celery cabbage, cauliflower and baby cabbage. This instrument was used to complete the qualitative and quantitative analysis of the TRFIS, so as to enhance the practical application of the detection method. This TRFIS possessed excellent linearity ranging from 0.25 mg kg−1 to 1.75 mg kg−1 for the detection of acetamiprid, and the limit of detection were 0.056–0.074 mg kg−1 in the different vegetable matrix. The platform combines the accuracy and portability of traditional test strips with the highly sensitive and efficient fluorescence intensity recognition function of detection equipment, which shows a great application prospect of multi-channel rapid detection of small molecule pollutants in the field.

Label-Free Electrochemical Aptasensor for Sensitive Detection of Malachite Green Based on AuNPs/MWCNTs@TiO2 Nanocomposites

This study proposes a label-free aptamer biosensor for the sensitive detection of malachite green(MG) using gold nanoparticles/multi-walled carbon nanotubes @ titanium dioxide(AuNPs/MWCNTs@TiO2). The nanocomposite provides a large surface area and good electrical conductivity, improving current transfer and acting as a platform for aptamer immobilization. The aptamer and the complementary chain(cDNA) are paired by base complementary to form the recognition element and fixed on the AuNPs by sulfhydryl group, which was modified on the cDNA. Since DNA is negatively charged, the redox probe in the electrolyte is less exposed to the electrode surface under the repulsion of the negative charge, resulting in a low-electrical signal level. When MG is present, the aptamer is detached from the cDNA and binds to MG, the DNA on the electrode surface is reduced, and the rejection of the redox probe is weakened, which leads to an enhanced electrical signal and enables the detection of MG concentration by measuring the change in the electrical signal. Under the best experimental conditions, the sensor demonstrates a good linear relationship for the detection of MG from 0.01 to 1000 ng/mL, the limit of detection (LOD)is 8.68 pg/mL. This sensor is stable, specific, and reproducible, allowing for the detection of various small-molecule pollutants by changing the aptamer, providing an effective method for detecting small-molecule pollutants.

Ultrasensitive and simultaneous monitoring of multiple small-molecule pollutants on an immunochromatographic strip with multilayered film-like fluorescent tags

A wide variety of small-molecule pollutants is harmful to human health, and their highly sensitive universal and rapid detection in complex environments remains a challenge. Herein, a multiplexed and ultrasensitive immunochromatographic strip (ICS) was developed for the universal analysis of three kinds of different pollutants based on multilayered fluorescent nanofilm-guided signal amplification. Flexible three-dimensional nanofilms (GO–MQD) with large surface areas, high quantum dot (QD) loading, superior luminescence, and good stability were synthesized through the electrostatic adsorption-mediated layer-by-layer assembly of three layers of small QDs onto two-dimensional graphene oxide (GO) nanosheets, modified with specific antibodies, and utilized as enhanced fluorescent tags in the ICS method for quantitative target detection. By combining the GO–MQD nanofilms and multiplexed ICS, the proposed assay can rapidly and sensitively detect aflatoxin B1, clenbuterol, and kanamycin in actual samples/environmental samples (pork extract, milk, river water, and lake water) with low detection limit (0.87, 2.04, and 0.81 pg/mL), fast testing time (15 min), good stability and high reproducibility (RSD < 8.71 %). The GO–MQD–ICS method developed here exhibits great potential to meet the demands of the on-site and practical detection of small-molecule pollutants.

Structure and function of ABC transporters in oceanic species.

The pollution of the world's oceans is a mounting problem that threatens the safety and well-being of our planet. Many marine organisms and ecosystems are exposed to pollutants with currently unknown toxicity and consequences. Membrane proteins that act as xenobiotic - foreign molecule - exporters are an important line of defense against pollutants in the organisms’ environment. One of the most ubiquitous membrane transporter proteins are the ATP binding cassette transporters (ABC). Found in organisms across all domains of life and in many tissue types, ABC transporters selectively export many different small molecules from within the cell, preventing the toxic accumulation of xenobiotics within the organism. Many of the small molecules exported by ABC transporters are pollutants. However, some small molecule pollutants may act as inhibitors of ABC transporters by interacting with their binding site and impeding transport. For example, a study conducted on one transporter - ABCB1 or P-glycoprotein - in mice found that transport activity was significantly reduced in the presence of DDD, DDE, and DDT. In the case of ABC transporters in oceanic species, many reagents found polluting the ocean may inhibit ABC transporters’ ability to export their wide range of substrates. This would lead to detrimental effects to the health of organisms in the oceanic food chain (e.g. yellowfin tuna) and to embryonic development (e.g. purple sea urchin). A thorough molecular and structural investigation of these transporters expressed in marine species is necessary to determine which chemicals inhibit transport. Using cryo-EM and x-ray crystallography, we intend to study the structure of the ABC transporter active site and the differences between activating and inhibiting molecular transport.

Steered photogenerated carrier separation via mixed-crystal-homojunction-mediated permanent negative charge electric field

The chelation between heavy metal ions and small organic molecules limits the prospects of traditional adsorption methods in dealing with the complex water environment, annihilating the non-negligible reutilization value of heavy metal ions. Photocatalysis enables the reduction of heavy metal ions and the mineralization of small molecule pollutants to be accomplished simultaneously, but the enhancement of photogenerated carrier separation efficiency is a technical challenge to be overcome. Herein, a photocatalyst loaded with mixed-crystal titanium dioxide (TiO2) on the surface of montmorillonite (MMT) was prepared, which is reported for the exceptional performance in the recovery of heavy metals Cr(VI) and the mineralization of organic pollutants Rh-B (10 ppm chelate in total) within 60 min. Scientific characterization and theoretical calculations demonstrate that the generation of oxygen vacancies in mixed-crystal homojunctions driven by a permanent negative charge electric field will maximize the adsorption capacity for H2O and O2. The MMT surface is converted into a reduction platform for Cr(VI), and anatase-TiO2 acts as an active site for Rh-B mineralization, under the synergistic effect of permanent negative charge electric field and mixed-crystal homojunctions. The Cr(III) obtained by photocatalytic reduction is collected by cation exchange between the interlayers of MMT with corresponding selective adsorption. This novel structural design concept provides a highly innovative orientation for the application of photocatalytic technology to the remediation of complex aqueous environments.

Universal and rapid detection of atrazine and bisphenol A using a reusable optical fiber chemiluminescent biosensor

Timely and accurately detection of small molecule pollutants is quite necessary to control environmental pollution and reduce harmfulness. Herein, a reusable optical fiber chemiluminescent biosensor (ROFC) was proposed for universal and rapid detection of two representative pollutants, pesticide atrazine (ATZ) and endocrine disruptor bisphenol A (BPA). The optical fiber modified with hapten-protein conjugates was regarded as both bio-probe and chemiluminescence signal transmission element, which effectively improved the light transmission efficiency and signal-to-noise ratio of the system. High-sensitive chemiluminescence signal detection is realized with a miniaturized ultrasensitive photodiode detector. Good regeneration performance of bio-probe can reduce detection cost and ensure detection reproducibility. Based on indirect competitive immunoassay principle, the chemiluminescence signal decreased with increasing pollutant concentration resulting from the less amount of antibody combined on the bio-probe surface. Under optimal conditions, the whole assay was achieved within 25 min with linear range of 1–100 μg/L and detection limits (LOD) for atrazine and BPA are 0.029 μg/L and 0.025 μg/L, respectively. The immunosensing optical fiber probe can be reused for 150 times at least without losing obvious bioactivity. The method was successfully applied to the detection of ATZ and BPA in three environmental samples, where recoveries between 93.4% and 116.6% were achieved. The ROFC biosensor provides a feasible platform for rapid detection of multiple small molecule pollutants in the environment.

Binding Between Antibiotics and Polystyrene Nanoparticles Examined by NMR.

Elucidating the interactions between plastic nanoparticles and small molecules is important to understanding these interactions as they occur in polluted waterways. For example, plastic that breaks down into micro- and nanoscale particles will interact with small molecule pollutants that are also present in contaminated waters. Other components of natural water, such as dissolved organic matter, will also influence these interactions. Here we use a collection of complementary NMR techniques to examine the binding between polystyrene nanoparticles and three common antibiotics, belonging to a class of molecules that are expected to be common in polluted water. Through examination of proton NMR signal intensity, relaxation times, saturation-transfer difference (STD) NMR, and competition STD-NMR, we find that the antibiotics have binding strengths in the order amoxicillin < metronidazole ≪ levofloxacin. Levofloxacin is able to compete for binding sites, preventing the other two antibiotics from binding. The presence of tannic acid disrupts the binding between levofloxacin and the polystyrene nanoparticles, but does not influence the binding between metronidazole and these nanoparticles.

Green-synthesized two-dimensional nanocarriers with photothermal capability for portable dual-readout immunochromatographic assay of clenbuterol

Immunochromatographic test strips (ITSs) have gained great practicability in point-of-care testing (POCT). Developing novel labels of ITSs with an additional signal to carry out quantitative analysis favors the development of sensitive POCT. Herein, tannic acid (TA) assisted stripping WS2 nanosheets (TA-WS2 NSs) based on colorimetric/photothermal ITS (TWITS) for clenbuterol (CLE) monitoring. After WS2 NSs loaded TA, it retained excellent photothermal performance and an intact surface structure for monoclonal antibody (mAb) labeling, at the same time, the stability of WS2 NSs and the activity of mAb were improved effectively. Satisfactory, the visual limit of detection (vLOD) of the photothermal signal was 0.3 ng/mL, which was enhanced 7-fold at least compared with traditional gold-based nanoparticles ITS (GNPs ITS). Importantly, this work can offer a new referenced method for other small molecule pollutants.

A review of spectroscopic probes constructed from aptamer-binding gold/silver nanoparticles or their dimers in environmental pollutants' detection.

The issue of environmental pollutant residues has gained wide public attention all along. Therefore, it is necessary to develop simple, rapid, economical, portable, and sensitive detection techniques, which have become the focus of research in the pollutants detection field. Spectroscopy is one of the most convenient, simple, rapid, and intuitive analytical tools that can provide accurate information, such as ultraviolet spectroscopy, fluorescence spectroscopy, Raman spectroscopy, plasmon resonance spectroscopy, etc. Gold nanoparticles, silver nanoparticles, and their dimers with unique optical properties are commonly used in the construction of spectroscopic probes. As a class of oligonucleotides that can recognize specific target molecules, aptamers also have a strong ability to recognize small-molecule pollutants. The application of aptamer-binding metal nanoparticles in biosensing detection presents significant advantages for instance high sensitivity, good selectivity, and rapid analysis. And many spectroscopic probes constructed by aptamer-binding gold nanoparticles, silver nanoparticles, or their dimers have been successfully demonstrated for detecting pollutants. This review summarizes the progress, advantages, and disadvantages of aptamer sensing techniques constructed by visual colorimetric, fluorescence, Raman, and plasmon resonance spectroscopic probes combining gold/silver nanoparticles or their dimers in the field of pollutants detection, and discusses the prospects and challenges for their future.

Investigating the Permeation Mechanism of Typical Phthalic Acid Esters (PAEs) and Membrane Response Using Molecular Dynamics Simulations.

Phthalic acid esters (PAEs) are typical environmental endocrine disrupters, interfering with the endocrine system of organisms at very low concentrations. The plasma membrane is the first barrier for organic pollutants to enter the organism, so membrane permeability is a key factor affecting their biological toxicity. In this study, based on computational approaches, we investigated the permeation and intramembrane aggregation of typical PAEs (dimethyl phthalate, DMP; dibutyl phthalate, DBP; di-2-ethyl hexyl phthalate, DEHP), as well as their effects on membrane properties, and related molecular mechanisms were uncovered. Our results suggested that PAEs could enter the membrane spontaneously, preferring the headgroup-acyl chain interface of the bilayer, and the longer the side chain (DEHP > DBP > DMP), the deeper the insertion. Compared with the shortest DMP, DEHP apparently increased membrane thickness, order, and rigidity, which might be due to its stronger hydrophobicity. Potential of means force (PMF) analysis revealed the presence of an energy barrier located at the water-membrane interface, with a maximum value of 2.14 kcal mol−1 obtained in the DEHP-system. Therefore, the difficulty of membrane insertion is also positively correlated with the side-chain length or hydrophobicity of PAE molecules. These findings will inspire our understanding of structure-activity relationship between PAEs and their effects on membrane properties, and provide a scientific basis for the formulation of environmental pollution standards and the prevention and control of small molecule pollutants.

Strategic design and fabrication of lightweight sesame ball-like hollow double-layer hybrid magnetic molecularly imprinted nanomaterials for the highly specific separation and recovery of tetracycline from milk

The synthesis of green and lightweight hollow double-layer magnetic adsorbents creates a roadmap for developing adsorbents for small molecule pollutants.

Universal fluorescence nanoprobes to enhance the sensitivity of immunochromatographic assay for detection of 17β-estradiol in milk

The pollution caused by estrogens in the environment and food has received increasing attention. It is still challenging for on-site immunochromatographic assay (ICA) detection of estrogens. The performance of the prepared probes plays a decisive role in the sensitivity and stability of the ICA system. The published probes usually directly couple the detection antibody to the label, ignoring the influence of the label on the activity of the antibody. In this study, 17β-estradiol (E2) was used as a model analyte for the ICA system. Two universal probes were constructed based on quantum dot nanobeads (QBs), recombinant protein A (SPA, from Staphylococcus aureus), and rabbit anti-mouse immunoglobulin G antibody (anti-IgG). The probes were prepared by coupling QBs with SPA, releasing anti-E2 monoclonal antibody (mAb), and maintaining its activity. The prepared universal probes can orient recognize the Fc region of mAb and fully expose its Fab region, improving the detection sensitivity of the ICA system. The free anti-E2 mAb and the universal probe (QBs@SPA or QBs@SPA@anti-IgG) were used as the detection antibodies and signal donors, respectively.The results show that the proposed ICA based on QBs@SPA and QBs@SPA@anti-IgG probes could detect E2 with IC50 of 8.83 and 0.93 ng/mL, respectively, within 15 min under optimal conditions. The recovery results of ICA based on QBs@SPA and QBs@SPA@anti-IgG probes showed good agreement with the findings of the high-performance liquid chromatography (HPLC) analysis for spiked samples. The developed ICA system based on universal probes was superior in terms of sensitivity, rapidity, and applicability, and held great promise for its implementation in detecting environmental and food small-molecule pollutants.