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result(s) for
"immunoaffinity chromatography"
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A New Abscisic Acid Conjugate, ABA-L-Glutamate, Determined in Different Plant Species by Combined Immunoaffinity Chromatography-Tandem Mass Spectrometry
by
Novák, Ondřej
,
Turečková, Veronika
,
Žukauskaitė, Asta
in
Abscisic acid
,
Agriculture
,
Amino acids
2024
Abscisic acid (ABA) is a phytohormone that occurs in plants at very low concentration (pmol/g fresh weight) and regulates multiple biological processes, including stomatal closure, seed germination, and responses to environmental stresses. In the present study, isolation of ABA, ABA glucosyl ester, and 11 ABA amino acid conjugates from minute quantities of plant tissue (less than 20 mg fresh weight) was achieved using a purification method based on the combination of an Oasis HLB column and an immunoaffinity sorbent. New monoclonal antibodies raised against (+)-
cis,trans
-ABA conjugated to BSA through its carboxyl group (C1) were characterised by enzyme-linked immunosorbent assay (ELISA) and used for immunoaffinity chromatography (IAC) gel preparation. The use of immunoaffinity purification significantly reduced matrix effects and increased the selectivity and sensitivity of subsequent UHPLC-MS/MS analysis. In addition to (+)-
cis,trans
-ABA and its glucosyl ester, a new abscisic acid conjugate, ABA-L-glutamate, was isolated by IAC and identified by tandem mass spectrometry in pea (
Pisum sativum
L.),
Lepidium sativum
L. and wheat (
Triticum aestivum
L.) seedlings. However, it was not found in 10-day-old seedlings of
Arabidopsis thaliana
or water-stressed tobacco (
Nicotiana tabacum
L.) leaves. Here, the identification of an ABA conjugate with glutamic acid in plants is described for the first time.
Journal Article
Advanced LC–MS-based methods to study the co-occurrence and metabolization of multiple mycotoxins in cereals and cereal-based food
2018
Liquid chromatography (LC) coupled with mass spectrometry (MS) is widely used for the determination of mycotoxins in cereals and cereal-based products. In addition to the regulated mycotoxins, for which official control is required, LC–MS is often used for the screening of a large range of mycotoxins and/or for the identification and characterization of novel metabolites. This review provides insight into the LC–MS methods used for the determination of co-occurring mycotoxins with special emphasis on multiple-analyte applications. The first part of the review is focused on targeted LC–MS approaches using cleanup methods such as solid-phase extraction and immunoaffinity chromatography, as well as on methods based on minimum cleanup (quick, easy, cheap, effective, rugged, and safe; QuEChERS) and dilute and shoot. The second part of the review deals with the untargeted determination of mycotoxins by LC coupled with high-resolution MS, which includes also metabolomics techniques to study the fate of mycotoxins in plants.
Journal Article
One-step time-resolved fluorescence microsphere immunochromatographic test strip for quantitative and simultaneous detection of DON and ZEN
2021
Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that contaminate a wide range of grains and crops. In this study, a one-step time-resolved single-channel immunochromatographic test strip based on europium ion polystyrene fluorescence microspheres was first developed for sensitive and quantitative detection of DON and ZEN. The concentration of the artificial antigen and the mass ratio of the monoclonal antibody to fluorescent microspheres for conjugation were optimized to simplify the sample addition process during immunochromatographic assay and improve the on-site detection efficiency. The limits of detection (LOD) of the single-channel immunochromatographic test strip for DON and ZEN detection were 0.17 and 0.54 μg/L, respectively. Meanwhile, the dual-channel immunochromatographic test strip was designed to simultaneously detect DON and ZEN, with LODs of 0.24 and 0.69 μg/L achieved for DON and ZEN, respectively. The developed test strips also yielded recovery results consistent with that obtained by LC-MS/MS for DON and ZEN detection in real samples of wheat and corn flour, confirming the practicability and reliability of the test strip. The developed immunochromatographic test strips realize quick and sensitive detection of DON and ZEN, exhibiting potential for broad applications in the point-of-care testing platform of multiple mycotoxins in agricultural products.
Journal Article
Sensing approaches on paper-based devices: a review
2013
Paper has been present in the world of analytical chemistry for centuries, but it seems that just a few years back it was rediscovered as a valuable substrate for sensors. We can easily list some of the countless advantages of this simple cellulosic substrate, including mechanical properties, three-dimensional fibrous structure, biocompatibility and biodegradability, easiness of production and modification, reasonable price, and availability all over the world. Those characteristics make paper a first-choice substrate for disposable sensors and integrated sensing platforms. Nowadays, numerous examples of paper-based sensors are being presented in the literature. This review describes some of the most prominent examples classifying them by type of detection: optical (colorimetric, fluorescence, surface-enhanced Raman spectroscopy, and transmittance methods) and electrochemical (voltammetric, potentiometric, and conductivity-based methods). We take a closer look at recent advances in immunoassays fabricated on paper, excluding simple lateral flow tests assembled on nitrocellulose. This review also summarizes the main advantages and disadvantages of the use of paper as a substrate for sensors, as well as its impact on their performance and application, presents a short history of paper in analytical chemistry, and discusses fabrication methods and available sources of paper.
Journal Article
Magnetic immunochromatographic test for histamine detection in wine
by
Socoliuc, Vlad
,
Moyano, Amanda
,
Martínez-García, José C
in
Beverages
,
Fermentation
,
Fermented food
2019
Histamine, a biogenic amine, is abundant in fermented foods and beverages, notably wine. A high intake of this monoamine may produce adverse reactions in humans, which may be severe in individuals with a reduced capacity to catabolise extrinsic histamine. Thus, control of histamine concentration during wine production and before distribution is advisable. Simple, rapid, point-of-use bioanalytical platforms are needed because traditional methods for the detection and quantification of histamine are expensive and time-consuming. This work applies the lateral flow immunoassay technique to histamine detection. Superparamagnetic particle labels, and an inductive sensor designed to read the test line in the immunoassay, enable magnetic quantification of the molecule. The system is calibrated with histamine standards in the interval of interest for wine production. A commercial optical strip reader is used for comparison measurements. The lateral flow system has a limit of detection of 1.2 and 1.5 mg/L for the inductive and optical readers, respectively. The capability of the inductive system for histamine quantification is demonstrated for wine samples at different processing points (at the end of alcoholic fermentation, at the end of malolactic fermentation, in freshly bottled wine, and in reserve wine). The results are validated by ultra-high-performance liquid chromatography.
Journal Article
Development of a p72 trimer–based colloidal gold strip for detection of antibodies against African swine fever virus
by
Ma, Hongfang
,
Lu, Qingxia
,
Qiao, Zixuan
in
African swine fever
,
African Swine Fever - diagnosis
,
African Swine Fever Virus
2022
African swine fever virus (ASFV) causes a highly contagious and often lethal swine viral disease, and leads to tremendous economic losses to the swine industry. Unfortunately, there are no vaccines and effective antiviral agents available to prevent and control ASFV outbreaks. Therefore, it is necessary to develop simple and rapid strategies to monitor ASFV-infected pigs to restrain its spread. In the current study, ASFV capsid protein p72 was expressed along with its chaperone pB602L to form trimers in human embryonic kidney 293 (HEK293) cells. The p72 trimers were subsequently labeled with colloidal gold to develop a immunochromatographic strip. The strip showed high specificity to ASFV-positive serum and no cross-reactivity to other swine virus positive sera. Importantly, the strip showed a higher sensitivity of detecting ASFV antibodies in both positive standard serum and clinical serum samples than a commercial enzyme-linked immunosorbent assay (ELISA) kit. Taken together, these results demonstrate the strip as a reliable diagnostic tool against ASFV infection, which will be appropriate for application in prevention and control of ASFV.
Key points
•
ASFV p72 trimers were successfully generated
.
•
A colloidal gold strip was developed based on ASFV p72 trimers
.
•
The strip is appropriate for detecting ASFV antibodies in the field
.
Journal Article
A novel dual-channel immunochromatographic strip using up-conversion nanoparticles for simultaneous detection of AFB1 and ZEN in maize
by
Shen, Yanghong
,
Zhu, Junli
,
Zhang, Jinzhi
in
Aflatoxin B1
,
Aflatoxin B1 - analysis
,
Aflatoxins
2023
Due to universal contamination and synergistic toxicity of multiple mycotoxins in foodstuff, reliable and high-throughput detection methods for multiple mycotoxins are urgently needed in corn products. In this study, a novel dual-channel immunochromatographic assay (ICA) based on improved up-conversion nanoparticles (IUCNPs) was developed for rapidly detecting aflatoxin B1 (AFB1) and zearalenone (ZEN). The synthesized IUCNPs doped by 30% Lu
3+
showed a larger size, more regular structure, and brighter fluorescence intensity than conventional UCNPs. The limits of detection (LODs) of single-channel ICA test strips for AFB1 and ZEN detection were 0.01 and 0.1 ng/mL, respectively. After the optimization, the dual-channel ICA of AFB1 and ZEN in 10 min was conducted, resulting in low detection limits of 0.025 and 0.1 ng/mL, respectively. Moreover, the built assay was revealed to be highly specific for six other food-contaminated mycotoxins, and exhibited excellent accuracy, with corresponding R
2
of 0.9931 and 0.9982 in calibration curves, respectively. Long-term storage experiments indicated that the dual-channel test strips had superior stability and precision. The LODs of AFB1 and ZEN in spiked maize were 0.025 and 0.25 μg/kg, demonstrating great sensitivity and matrix tolerance. Furthermore, the IUNCP-ICA was validated by high-performance liquid chromatography (HPLC) analyses, and a satisfactory consistency was obtained in 15 natural maize samples. Thus, the IUCNPs-ICA proposed in this work realized rapid and sensitive detection of AFB1 and ZEN, providing broad application potential in on-site screening for multiple mycotoxins in agricultural products.
Graphical abstract
Journal Article
Affinity monolith chromatography: a review of principles and recent analytical applications
by
Pfaunmiller, Erika L.
,
Paulemond, Marie Laura
,
Dupper, Courtney M.
in
Affinity
,
Analytical Chemistry
,
Analytical instruments
2013
Affinity monolith chromatography (AMC) is a type of liquid chromatography that uses a monolithic support and a biologically related binding agent as a stationary phase. AMC is a powerful method for the selective separation, analysis, or study of specific target compounds in a sample. This review discusses the basic principles of AMC and recent developments and applications of this method, with particular emphasis being given to work that has appeared in the last 5 years. Various materials that have been used to prepare columns for AMC are examined, including organic monoliths, silica monoliths, agarose monoliths, and cryogels. These supports have been used in AMC for formats that have ranged from traditional columns to disks, microcolumns, and capillaries. Many binding agents have also been employed in AMC, such as antibodies, enzymes, proteins, lectins, immobilized metal ions, and dyes. Some applications that have been reported with these binding agents in AMC are bioaffinity chromatography, immunoaffinity chromatography or immunoextraction, immobilized-metal-ion affinity chromatography, dye–ligand affinity chromatography, chiral separations, and biointeraction studies. Examples are presented from fields that include analytical chemistry, pharmaceutical analysis, clinical testing, and biotechnology. Current trends and possible directions in AMC are also discussed.
Journal Article
Aflatoxin detection technologies: recent advances and future prospects
by
Yao, Zhiliang
,
Liu, Minhua
,
Liu, Shenqi
in
Aflatoxins
,
Aflatoxins - analysis
,
Aquatic Pollution
2023
Aflatoxins have posed serious threat to food safety and human health. Therefore, it is important to detect aflatoxins in samples rapidly and accurately. In this review, various technologies to detect aflatoxins in food are discussed, including conventional ones such as thin-layer chromatography (TLC), high performance liquid chromatography (HPLC), enzyme linked immunosorbent assay (ELISA), colloidal gold immunochromatographic assay (GICA), radioimmunoassay (RIA), fluorescence spectroscopy (FS), as well as emerging ones (e.g., biosensors, molecular imprinting technology, surface plasmon resonance). Critical challenges of these technologies include high cost, complex processing procedures and long processing time, low stability, low repeatability, low accuracy, poor portability, and so on. Critical discussion is provided on the trade-off relationship between detection speed and detection accuracy, as well as the application scenario and sustainability of different technologies. Especially, the prospect of combining different technologies is discussed. Future research is necessary to develop more convenient, more accurate, faster, and cost-effective technologies to detect aflatoxins.
Graphical abstract
Journal Article
Development and application of a colloidal-gold dual immunochromatography strip for detecting African swine fever virus antibodies
by
Wan, Ying
,
Song, Rui
,
Zhou, Gaijing
in
African swine fever
,
African Swine Fever - diagnosis
,
African Swine Fever Virus
2022
African swine fever (ASF) is an acute and highly contagious infectious disease caused by the African swine fever virus (ASFV). Currently, there is no vaccine against ASF worldwide, and no effective treatment measures are available. For this reason, developing a simple, rapid, specific, and sensitive serological detection method for ASFV antibodies is crucial for the prevention and control of ASF. In this study, a 1:1 mixture of gold-labeled p30 and p72 probes was used as the gold-labeled antigen. The p30 and p72 proteins and their monoclonal antibodies were coated on a nitrocellulose membrane (NC) as a test (T) line and control (C) line, respectively. A colloidal-gold dual immunochromatography strip (ICS) for ASFV p30 and p72 protein antibodies was established. The results showed that the colloidal-gold dual ICS could specifically detect ASFV antibodies within 5–10 min. There was no cross-reaction after testing healthy pig serum; porcine reproductive and respiratory syndrome virus (PRRSV), foot-and-mouth disease type A virus (FMDV-A), foot-and-mouth disease type O virus (FMDV-O), porcine circovirus type 2 (PCV-2), and classical swine fever virus (CSFV) positive sera. A positive result was obtained only for the positive control P1. The sensitivity of the test strips was 1:256, which was equivalent to that of commercially ELISA kits. Their coincidence rate with the two commercial ASFV ELISA antibodies detection kits was higher than 98%. The test strips were stably stored at 18–25 °C and 4 °C for 4 and 6 months, respectively. The colloidal-gold dual ICS prepared in this study had high sensitivity and specificity and were characterized by rapid detection, simple operation, and easy interpretation of results. Therefore, they are of great significance to diagnose, prevent, and control African swine fever.
Key points
•
We establish an antibody detection that is quick and can monitor an ASF infection.
•
We observe changes in two protein antibodies to dynamically monitor ASF infection.
•
We use diversified detection on a single test strip to detect both antibodies.
Journal Article