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Keywords = immuno-magnetic adsorbent

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12 pages, 1594 KB  
Article
pH-Regulated Strategy and Mechanism of Antibody Orientation on Magnetic Beads for Improving Capture Performance of Staphylococcus Species
by Fuying Kang, Yin Yang, Jingwen Li, Erning Chen, Tian Hong, Lulu Zhao and Meihong Du
Foods 2022, 11(22), 3599; https://doi.org/10.3390/foods11223599 - 11 Nov 2022
Cited by 6 | Viewed by 4429
Abstract
Immunomagnetic beads (IMBs) have been widely used to capture and isolate target pathogens from complex food samples. The orientation of the antibody immobilized on the surface of magnetic beads (MBs) is closely related to the effective recognition with an antigen. We put forward [...] Read more.
Immunomagnetic beads (IMBs) have been widely used to capture and isolate target pathogens from complex food samples. The orientation of the antibody immobilized on the surface of magnetic beads (MBs) is closely related to the effective recognition with an antigen. We put forward an available strategy to orient the antibody on the surface of MBs by changing the charged amino group ratio of the reactive amino groups at optimal pH value. Quantum dots labeling antigen assay, antigen-binding fragment (Fab) accessibility assay and lysine mimicking were used for the first time to skillfully illustrate the antibody orientation mechanism. This revealed that the positively charged ε-NH2 group of lysine on the Fc relative to the uncharged amino terminus on Fab was preferentially adsorbed on the surface of MBs with a negatively charged group at pH 8.0, resulting in antigen binding sites of antibody fully exposed. This study contributes to the understanding of the antibody orientation on the surface of MBs and the potential application of IMBs in the separation and detection of pathogenic bacteria in food samples. Full article
(This article belongs to the Special Issue Application of Immunoassay Technology in Food Inspection)
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15 pages, 3274 KB  
Article
An Immuno-Separated Assay for Ochratoxin Detection Coupled with a Nano-Affinity Cleaning-Up for LC-Confirmation
by Jie-Biao Guo, Jin-Sheng Cheng, Tai-Long Wei, Fan-Min Wu, Gui-Hong Tang and Qing-Hua He
Foods 2022, 11(8), 1155; https://doi.org/10.3390/foods11081155 - 15 Apr 2022
Cited by 3 | Viewed by 2796
Abstract
An immuno-separated assay for ochratoxin A detection coupled with a nano-affinity cleaning up for LC-confirmation was developed. Firstly, ochratoxin A was modified to quantum dot beads for immuno-fluorescent reporters. Secondly, Fe3O4 magnetic nanoparticles were conjugated with protein G for immuno-magnetic [...] Read more.
An immuno-separated assay for ochratoxin A detection coupled with a nano-affinity cleaning up for LC-confirmation was developed. Firstly, ochratoxin A was modified to quantum dot beads for immuno-fluorescent reporters. Secondly, Fe3O4 magnetic nanoparticles were conjugated with protein G for immuno-magnetic adsorbents. The immuno-separation of fluorescent reporters by magnetic adsorbents could be completed by ochratoxin A, so the fluorescent reporters released from the immune complex indicate a linear correlation with the concentration of ochratoxin A. Furthermore, the immuno-separated ochratoxin A can be eluted from magnetic adsorbent for LC-conformation. The optimized assay showed results as follows: the quantitative range of the immuno-separated assay was 0.03–100 ng mL−1 of ochratoxin A. The recoveries for spiked samples ranged from 78.2% to 91.4%, with the relative standard deviation (RSD) being 11.9%~15.3%. Statistical analysis indicated no significant difference between the HPLC-FLD results based on commercial affinity column and by nano-affinity cleaning up. Full article
(This article belongs to the Topic Future Food Analysis and Detection)
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