Dataset Information

PH-Regulated Strategy and Mechanism of Antibody Orientation on Magnetic Beads for Improving Capture Performance of Staphylococcus Species.

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 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.

SUBMITTER: Kang F

PROVIDER: S-EPMC9689862 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Publications

pH-Regulated Strategy and Mechanism of Antibody Orientation on Magnetic Beads for Improving Capture Performance of <i>Staphylococcus</i> Species.

Kang Fuying F Yang Yin Y Li Jingwen J Chen Erning E Hong Tian T Zhao Lulu L Du Meihong M

Foods (Basel, Switzerland) 20221111 22

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 ...[more]

PMID: 36429188

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Project description:Diabetes is a chronic metabolic disease which leads to high glucose levels in the blood, with severe consequences for human health. Due to the worldwide appeal for the reduction in calorie intake, this study presents the development of a nanomaterial able to capture sucrose selectively, thus providing a tool to remove naturally occurring sucrose from food, such as fruit juices, producing low-calorie juices for consumption. Magnetite nanoparticles (Fe3O4 NPs) coated with an inert material (SiO2) and functionalised with the enzyme invertase were designed to remove sucrose from solutions. Fe3O4 NPs were synthesised using the co-precipitation method, whereas the coating with a silica shell was done by the Stöber method. Its physicochemical characteristics were determined, with excellent stability over time. On the other hand, the invertase enzyme was extracted from dry Baker's yeast, purified and immobilised on the surface of the silica-coated Fe3O4 NPs. pH-triggered sucrose capture occurred at pH 3.0 once invertase with protonated catalytic residues was able just to bind with sucrose in a highly selective way. After a short, 1 min interaction, approximately 13.5 mmol L-1 of sucrose was captured per gram of nanomaterial and removed with the use of an external permanent magnet. The complex sucrose/nanomaterial was washed, and the released sucrose was put into buffered solution (pH = 4.8), where it underwent hydrolysis to yield inverted sugar. On the other side, sucrose-free nanomaterial was reused with no loss of enzymatic capability to capture sucrose at pH = 3.0 and maintained the invertase activity at pH 4.8 in ten consecutive rounds of re-use. As sucrose was recovered in the form of inverted sugar, not just low sugar beverage could be obtained, but also a high valued market product. Thus, the developed technology allows for the commercialisation of low-calorie food, offering healthier options to consumers and helping to fight diabetes and obesity.

Dataset Information

PH-Regulated Strategy and Mechanism of Antibody Orientation on Magnetic Beads for Improving Capture Performance of Staphylococcus Species.

Publications

pH-Regulated Strategy and Mechanism of Antibody Orientation on Magnetic Beads for Improving Capture Performance of <i>Staphylococcus</i> Species.

Similar Datasets

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