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Multicolor immunoassay is a powerful tool for rapid analysis without the use of bulky instruments owing to various color conversions, which is suitable for on-site visual analysis for pesticides. Herein, this study developed a multicolor immunoassay for the rapid detection of isocarbophos. After competitive immunoassay, the secondary antibody (GAM-ALP) catalyzed ascorbyl-2-phosphate (AAP) into ascorbic acid (AA). The AA can reduce K[Fe(CN)] into K[Fe(CN)]. The latter can react with Fe to form Prussian blue; otherwise, the orange AAP-Fe complex was generated. Therefore, the multicolor immunoassay achieved a color conversion of orange-green-blue in response to isocarbophos, allowing for rapid semiquantitative analysis by the naked eye. After parameter optimization, the multicolor immunoassay was developed depending on the ratiometric absorbance between the Prussian blue and AAP-Fe complex. Moreover, a smartphone was used to measure the RGB value of the color conversion for the development of portable visual, quantitative analysis. Both the absorbance-based and RGB-based multicolor immunoassays showed good accuracy and practicability in the recovery test. This study provided a common approach for the development of dual-readout multicolor immunoassay, which can be used for on-site rapid screening by quantitative or visual semiquantitative analysis.
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http://dx.doi.org/10.3390/foods13244057 | DOI Listing |
Biochemistry (Mosc)
May 2025
Division of Immunobiology and Biomedicine, Scientific Center for Genetics and Life Sciences, , Sirius University of Science and Technology, Sirius, 354340, Russia.
Bacterial β-galactosidase (LacZ) has been widely used as a reporter for the development of mice models to study gene expression or for control of conditional gene deletion. However, high level of endogenous β-galactosidase expression, and low sensitivity of existing substrates for detection of transgenic LacZ activity limited this reporter application for live cell analysis. To overcome this limitation, we evaluated performance of the intracellularly immobilizable fluorescent probe SPiDER-βGal to detect LacZ in major blood cell populations of the reporter mice using multicolor flow cytometry.
View Article and Find Full Text PDFCancer Cell Int
March 2025
Department of Gynecology and Obstetrics, University Medical Center Regensburg, Regensburg, Germany.
Background: The immune checkpoint targeting is nowadays an integral part of cancer therapies. However, only a minority of patients experience long-term benefits. Thus, the identification of predictive biomarkers contributing to therapy response is urgently needed.
View Article and Find Full Text PDFAnalyst
April 2025
Department of Neurological Rehabilitation, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou 325027, P.R. China.
Sepsis is a systemic inflammatory response syndrome caused by infection, requiring the joint detection of multiplex biomarkers for specific diagnosis. Here, we present a chemiluminescence enzyme immunoassay based on microfluidic magnetic droplets for multiplex sepsis biomarker screening. The droplet-based chemiluminescence enzyme immunoassay (CLIA) technology utilizes multicolor-encoded microspheres to distinguish biomarkers and mesoporous silica-loaded enzymes for signal amplification and catalytic fluorescent substrates.
View Article and Find Full Text PDFFoods
December 2024
Key Laboratory of Tropical Fruit and Vegetable Cold-Chain of Hainan Province, Institute of Agro-Products of Processing and Design, Hainan Academy of Agricultural Sciences, Haikou 571100, China.
Multicolor immunoassay is a powerful tool for rapid analysis without the use of bulky instruments owing to various color conversions, which is suitable for on-site visual analysis for pesticides. Herein, this study developed a multicolor immunoassay for the rapid detection of isocarbophos. After competitive immunoassay, the secondary antibody (GAM-ALP) catalyzed ascorbyl-2-phosphate (AAP) into ascorbic acid (AA).
View Article and Find Full Text PDFTalanta
May 2025
Marshall Laboratory of Biomedical Engineering, Shenzhen Key Laboratory for Nano-Biosensing Technology, Department of Biomedical Engineering, Medical School, Shenzhen University, Shenzhen, Guangdong, 518060, PR China. Electronic address:
Colorimetric enzyme-linked immunosorbent assays (CELISAs) have long been used for protein biomarker detection in diagnostics. Unfortunately, as confined by the monochromatic nature of detection signals and the limited catalytic activity of enzymes, CELISAs suffer from poor visual resolution and low sensitivity, hindering their effectiveness for early diagnostics in resource-limited settings. Herein, we report an ultrasensitive, high-visual-resolution CELISA (named PE-TSA-AuAg Cage-CELISA) that combines kinetically controlled growth of Ag in AuAg nanocages with poly-enzyme-boosted tyramide signal amplification (PE-TSA), enabling visual semiquantitative detection of protein biomarkers at attomolar levels with the naked eye.
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