Pocsfalvi Gabriella, Schlosser Gitta
Institute of Protein Biochemistry, National Research Council, Naples, Italy.
Methods Mol Biol. 2011;739:3-12. doi: 10.1007/978-1-61779-102-4_1.
Bacterial protein toxins are involved in a number of infectious and foodborne diseases and are considered as potential biological warfare agents as well. Their sensitive multiplex detection in complex environmental, food, and biological samples are an important although challenging task. Solid-phase immunoaffinity capture provides an efficient way to enrich and purify a wide range of proteins from complex mixtures. We have shown that staphylococcal enterotoxins, for example, can be efficiently enriched by means of magnetic immunocapture using antibody functionalized paramagnetic beads. The method was successfully interfaced by the on-beads and off-beads detection using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry at the protein level and by the off-beads nano-electrospray ionization-MS/MS detection at the enzyme digests level, enabling thus the unambiguous identification of the toxin. The method is applicable to any bacterial toxin to which an antibody is available.
细菌蛋白毒素与多种传染病和食源性疾病有关，同时也被视为潜在的生物战剂。在复杂的环境、食品和生物样品中对其进行灵敏的多重检测是一项重要但具有挑战性的任务。固相免疫亲和捕获提供了一种从复杂混合物中富集和纯化多种蛋白质的有效方法。例如，我们已经表明，使用抗体功能化的顺磁性珠子通过磁性免疫捕获可以有效地富集葡萄球菌肠毒素。该方法通过在蛋白质水平上使用基质辅助激光解吸/电离飞行时间质谱进行珠上和珠外检测，以及在酶消化水平上进行珠外纳米电喷雾电离-MS/MS检测成功实现联用，从而能够明确鉴定毒素。该方法适用于任何可获得抗体的细菌毒素。
