Antibody-integrated and functionalized graphite-encapsulated magnetic beads, produced using ammonia gas plasma technology, for capturing Salmonella
Antibody-integrated and functionalized graphite-encapsulated magnetic beads, produced using ammonia gas plasma technology, for capturing Salmonella
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使用氨气等离子体技术生产的抗体集成和功能化石墨封装磁珠，用于捕获沙门氏菌
DOI：
10.1016/j.bmcl.2015.01.031
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
and Masaaki Nagatsu
中科院分区：
文献类型：
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作者：
Akikazu Sakudo;Han Chou;and Masaaki Nagatsu
沙门氏菌是食源性感染的唯一和最重要的病原体，特别是涉及鸡蛋，牛奶和肉类等食物。为了预防感染，需要一个可靠的监测系统，可以快速，灵敏地检测沙门氏菌。在这里，我们描述了抗体集成的磁珠功能化的一种新的策略，使用氨气等离子体的发展。通过射频（RF）电源产生的氨等离子体被允许与石墨封装的磁珠的表面反应，导致氨基的引入。然后通过N-琥珀酰亚胺基3-（2-吡啶基二硫代）丙酸酯（SPDP）将抗沙门氏菌抗体通过存在于蛋白质表面上的硫化物基团锚定到磁珠的氨基。这些磁珠用于捕获沙门氏菌的潜在有用性被检查如下。将微珠与沙门氏菌蛋白液体培养基一起孵育，然后通过施加磁场从上清液中分离。彻底洗涤后，通过免疫层析、聚合酶链反应和直接培养测定证实沙门氏菌吸附到珠上。我们的研究结果表明，使用抗体整合的磁珠捕获和浓缩沙门氏菌比商业Dynabeads®抗沙门氏菌更有效，后者通常用于从液体培养物中浓缩沙门氏菌。我们相信这种新的珠技术将有助于提高沙门氏菌的检测。
Salmonellaspp. is the single and most important causative agent of foodborne infections, especially involving foods such as eggs, milk and meat. To prevent infection, a reliable surveillance system is required that can quickly and sensitively detectSalmonella. Here, we describe the development of antibody-integrated magnetic beads that are functionalized by a novel strategy using ammonia gas plasma. Ammonia plasma, produced by a radio frequency (RF) power supply, was allowed to react with the surface of graphite-encapsulated magnetic beads, resulting in the introduction of amino groups. An anti-Salmonellaantibody was then anchored by sulfide groups present on the protein surface to the amino groups of the magnetic beads viaN-succinimidyl 3-(2-pyridyldithio) propionate (SPDP). The potential usefulness of these magnetic beads for capturingSalmonellawas examined as follows. The beads were incubated withSalmonellain liquid medium and then separated from the supernatant by applying a magnetic field. After thorough washing, adsorption ofSalmonellato the beads was confirmed by immunochromatography, polymerase chain reaction and a direct culture assay. Our findings indicate that the capture and concentration ofSalmonellausing the antibody-integrated magnetic beads was more efficient than commercial Dynabeads® anti-Salmonella, which are conventionally used for concentratingSalmonellafrom liquid cultures. We believe this novel bead technology will contribute to the enhanced detection ofSalmonella.