Preparation of arsenic(III) monoclonal antibodies and preliminary evaluation of a novel silver-coated gold nanorod SERS immunoassay strip construction.

Heavy metal pollution has become a growing concern in industrial, agricultural, and manufacturing processes, posing a significant threat to human health. Among these heavy metals, arsenic (As) is highly toxic and shares similar chemical properties and environmental behavior with other heavy metals. As(III) is particularly toxic compared to other forms of arsenic. Therefore, it is essential to develop a real-time, rapid, and sensitive method for the determination of As(III). In this study, we employed a unique bifunctional chelator, 1-(4-isothiocyanobenzyl)-ethylenediamine N,N,N',N'-tetraacetic acid (ITCBE), to prepare a complete antigen. Through a series of tests including balb/c mouse immunization, cell fusion (mouse L2041 spleen cells with mouse myeloma cells SP2/0), and subcloning, we generated four monoclonal cell lines (1C1, 2C2, 3A9, and 4A11). These cell lines demonstrated high purity, high affinity, and IC50 values of less than 50 µg mL-1. Monoclonal antibody 4A11, which exhibited a strong Raman signal, was selected as the probe, and Au@Ag 200 was utilized as the surface-enhanced Raman scattering (SERS) substrate for the preliminary establishment of SERS immunochromatographic test strips. The sensitivity of the SERS immunochromatographic test strips, measured through Raman signal detection, showed a significant improvement compared to the SERS immunochromatographic test strips analyzed by colorimetry (LOD = 49.43 µg mL-1 and LDR = 5.32-81.31 µg mL-1). The SERS immunochromatographic test strips achieved a LOD of 7.62 µg mL-1 and an LDR of 12.66-71.84 µg mL-1. This study presents innovative methodologies for the rapid detection of As(III) using SERS immunochromatographic test strips.

High-Performance Au@Ag Nanorods Substrate for SERS Detection of Malachite Green in Aquatic Products.

In order to improve the detection performance of surface-enhanced Raman scattering (SERS), a low-cost Au@Ag nanorods (Au@Ag NRs) substrate with a good SERS enhancement effect was developed and applied to the detection of malachite green (MG) in aquaculture water and crayfish. By comparing the SERS signal enhancement effect of five kinds of Au@Ag NRs substrates with different silver layer thickness on 4-mercaptobenzoic acid (4-MBA) solution, it was found that the substrate prepared with 100 µL AgNO3 had the smallest aspect ratio (3.27) and the thickest Ag layer (4.1 nm). However, it showed a good signal enhancement effect, and achieved a detection of 4-MBA as low as 1 × 10-11 M, which was 8.7 times higher than that of the AuNRs substrate. In addition, the Au@Ag NRs substrate developed in this study was used for SRES detection of MG in crayfish; its detection limit was 1.58 × 10-9 M. The developed Au@Ag NRs sensor had the advantages of stable SERS signal, uniform size and low cost, which provided a new tool for SERS signal enhancement and highly sensitive SERS detection method development.

Preparation, Identification and Preliminary Application of the Fenvalerate Monoclonal Antibody in Six Kinds of Dark Tea.

Fenvalerate has the advantages of a broad insecticidal spectrum, high efficiency, low toxicity and low cost, and it is widely used in agriculture, especially in tea, resulting in the accumulation of fenvalerate residues in tea and the environment, posing a serious threat to human health. Therefore, the timely monitoring of fenvalerate residue dynamics is vital for ensuring the health of humans and the ecological environment, and it is necessary for establishing a fast, reliable, accurate and on-site method for detecting fenvalerate residues. Based on the methods of immunology, biochemistry and molecular biology, mammalian spleen cells, myeloma cells and mice were used as experimental materials to establish a rapid detection method of an enzyme-linked immunosorbent assay to detect the residues of fenvalerate in dark tea. Three cell lines-1B6, 2A11 and 5G2-that can stably secrete fenvalerate antibodies were obtained by McAb technology, and their sensitivities (IC50) were 36.6 ng/mL, 24.3 ng/mL and 21.7 ng/mL, respectively. The cross-reaction rates of the pyrethroid structural analogs were all below 0.6%. Six dark teas were used to detect the practical application of fenvalerate monoclonal antibodies. The sensitivity IC50 of the anti-fenvalerate McAb in PBS with 30% methanol is 29.12 ng/mL. Furthermore, a latex microsphere immunochromatographic test strip with an LOD of 10.0 ng/mL and an LDR of 18.9-357 ng/mL was preliminarily developed. A specific and sensitive monoclonal antibody for fenvalerate was successfully prepared and applied to detect fenvalerate in dark teas (Pu'er tea, Liupao tea, Fu Brick tea, Qingzhuan tea, Enshi dark tea and selenium-enriched Enshi dark tea). A latex microsphere immunochromatographic test strip was developed for the preparation of rapid detection test strips of fenvalerate.

MTM1 displays a new function in the regulation of nickel resistance in Saccharomyces cerevisiae.

Nickel (Ni) is an essential yet toxic trace element. Although a cofactor for many metalloenzymes, nickel function and metabolism is not fully explored in eukaryotes. Molecular biology and metallomic methods were utilized to explore the new physiological functions of nickel in Saccharomyces cerevisiae. Here we showed that MTM1 knockout cells displayed much stronger nickel tolerance than wild-type cells and mitochondrial accumulations of Ni and Fe of mtm1Δ cells dramatically decreased compared to wild-type cells when exposed to excess nickel. Superoxide dismutase 2 (Sod2p) activity in mtm1Δ cells was severely attenuated and restored through Ni supplementation in media or total protein. SOD2 mRNA level of mtm1Δ cells was significantly higher than that in the wild-type strain but was decreased by Ni supplementation. MTM1 knockout afforded resistance to excess nickel mediated through reactive oxygen species levels. Meanwhile, additional Ni showed no significant effect on the localization of Mtm1p. Our study reveals the MTM1 gene plays an important role in nickel homeostasis and identifies a novel function of nickel in promoting Sod2p activity in yeast cells.

Immunosensor of Nitrofuran Antibiotics and Their Metabolites in Animal-Derived Foods: A Review.

Nitrofuran antibiotics have been widely used in the prevention and treatment of animal diseases due to the bactericidal effect. However, the residual and accumulation of their metabolites in vivo can pose serious health hazards to both humans and animals. Although their usage in feeding and process of food-derived animals have been banned in many countries, their metabolic residues are still frequently detected in materials and products of animal-derived food. Many sensitive and effective detection methods have been developed to deal with the problem. In this work, we summarized various immunological methods for the detection of four nitrofuran metabolites based on different types of detection principles and signal molecules. Furthermore, the development trend of detection technology in animal-derived food is prospected.

Assessment of Contents and Health Impacts of Four Metals in Chongming Asparagus-Geographical and Seasonal Aspects.

In this paper, the contents of four typical metals (Pb, Cd, Hg, and As) in asparagus, water, and soil from Chongming Island were quantitatively determined by inductively coupled plasma mass spectrometry (ICP-MS). The contents of these metals in asparagus showed a common rule of Pb > As > Cd > Hg in different harvest seasons and regions. Significant seasonal differences were found in the contents by difference analysis, but no obvious regional differences were observed. Furthermore, the asparagus did not accumulate these four metals from the soil in Chongming Island by the assessment of bio-concentration factor. The asparagus was proved safe by the analysis of single-factor pollution index and Nemerow pollution index. Through combining the analysis of the above indexes and the geological accumulation index, we found that 51.62% of soil samples were mildly polluted by cadmium. The results of health risk analysis showed that the risk value of children was higher than that of adults under oral exposure, but the four metals in asparagus possessed no obvious risk to health. The above assessments illustrate that the daily consumption of asparagus in Chongming Island will not cause potential health impacts. It is of benefit to ensure the quality and economic interests of asparagus planting in Chongming Island through the investigation of this study.

Immunoassay based on Au-Ag bimetallic nanoclusters for colorimetric/fluorescent double biosensing of dicofol.

The high toxicity of dicofol (DICO) to nontarget organisms has resulted in the contamination of food materials and caused a threat to human health. Developing a rapid and sensitive detection method of DICO in food samples is essential and still pursued. Fluorescent nanomaterials have been widely applied in biosensors to improve the sensitivity of detection. Herein, glutathione-capped Au-Ag bimetallic nanoclusters (Au-Ag NCs) exhibited the outstanding fluorescence characteristic with the average fluorescence lifetime of 1971.08 ns and photoluminescence quantum yield of 9.84% when the molar ratio of Au to Ag was 5:1. Polyethyleneimine modified gold nanoparticles (PEI-Au NPs) with the positive charge were prepared to generate a strong colorimetric signal. A dual-model colorimetric/fluorescent immune probe based on the Au-Ag NCs and PEI-Au NPs was successfully constructed by electrostatic force, and could be applied in both ic-ELISA and LFIA methods for rapid and ultrasensitive detection of DICO. In the ic-ELISA method, the introduction of fluorescence signal significantly increased the sensitivity of detection with the limit of detection (LOD) of 0.62 ng/mL and exhibited an excellent linear relationship within the range of 1.36 ng/mL-19.92 ng/mL. In the LFIA method, the fluorescence signal of Au-Ag NCs was accumulated on the test line and control line for the fluorescence model detection with a quantitative LOD at the level of 1.59 ng/mL. Such a dual-model colorimetric/fluorescent immunoassay serves as a promising candidate to develop new approaches in field detection.

Novel monoclonal antibody-sandwich immunochromatographic assay based on Fe3O4/Au nanoparticles for rapid detection of fish allergen parvalbumin.

In this study, a rapid sandwich immunochromatographic assay (ICA) was developed to detect parvalbumin (PV). Firstly, two optimum primary monoclonal antibody (mAb) against PV had been screened out: mAb1 was used as the capture antibody, and mAb2 conjugated to Fe3O4/Au nanoparticles (Fe3O4/AuNPs) that served as a detection reagent. Using this pair of mAbs, a sandwich ICA strip based on Fe3O4/AuNPs was developed. The results showed that the color intensity of test line positively correlated with the PV concentration in the standard or spiked sample. The limit of detection for qualitative (LOD) and quantitative detection (LOQ) were 2 ng/mL and 0.691 ng/mL, respectively. Besides, the detection time of this ICA strip was within 15 min. The recovery rates ranged from 104.0% to 117.4%, within an acceptable level (80-120%). Moreover, the developed assay also showed high cross reaction in different fish species. These results demonstrated that the established test strip has the potential to be used as a rapid screening tool for large scale determination of PV in foodstuffs.

Novel Latex Microsphere Immunochromatographic Assay for Rapid Detection of Cadmium Ion in Asparagus.

Recently, concerns about heavy metal cadmium ion (Cd2+) residue in asparagus have been frequently reported, and there is an urgent need to develop an effective, sensitive, and rapid detection method for Cd2+. In this study, we innovatively combined molecular microbiology to carry out the comparative screening of Cd2+ chelators in a green, efficient, and specific way. The knock-out putative copper-transporter gene (pca1Δ) yeast strain with high sensitivity to Cd2+ was first used to screen the Cd2+ chelator, and the optimum chelator 1-(4-Isothiocyanatobenzyl)ethylenediamine-N,N,N,N'-tetraacetic acid (ITCBE) was obtained. Additionally, a rapid latex microsphere immunochromatographic assay (LMIA) was developed, based on the obtained monoclonal antibody (mAb) with high specificity and high affinity (affinity constant Ka = 1.83 × 1010 L/mol), to detect Cd2+ in asparagus. The 50% inhibitive concentration (IC50) of test strip was measured to be 0.2 ng/mL, and the limit of detection (IC10) for qualitative (LOD, for visual observation) and quantitative detection (LOQ, for data simulation) of the test strip was 2 ng/mL and 0.054 ng/mL, respectively. In all, the developed mAb-based LMIA shows a great potential for monitoring Cd2+ in asparagus, even in vegetable samples.

Multifunctional magnetoplasmonic nanoparticle assemblies for cancer therapy and diagnostics (theranostics).

In this work, we describe the preparation and biomedical functionalities of complex nanoparticle assemblies with magnetoplasmonic properties suitable for simultaneous cancer therapy and diagnostics (theranostics). Most commonly magnetoplasmonic nanostructures are made by careful adaptation of metal reduction protocols which is both tedious and restrictive. Here we apply the strategy of nanoscale assemblies to prepare such systems from individual building blocks. The prepared superstructures are based on magnetic Fe(3) O(4) nanoparticles encapsulated in silica shell representing the magnetic module. The cores are surrounded in a corona-like fashion by gold nanoparticles representing the plasmonic module. As additional functionality they were also coated by poly(ethyleneglycol) chains as a cloaking agent to extend the blood circulation time. The preparation is exceptionally simple and allows one to vary the contribution of each function. Both modules can carry drugs and, in this study, they were loaded with the potential anticancer drug curcumin. A comprehensive set of microscopy, spectroscopy and biochemical methods were applied to characterize both imaging and therapeutic function of the nanoparticle assemblies against leukemia HL-60 cells. High contrast magnetic resonance images and high apoptosis rates demonstrate the success of assembly approach for the preparation of magnetoplasmonic nanoparticles. This technology allows one to easily "dial in" the functionalities in the clinical setting for personalized theranostic regiments.