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A novel mycotoxin purification system using magnetic nanoparticles for the recovery of aflatoxin B1 and zearalenone from feed

机译:使用磁性纳米颗粒从饲料中回收黄曲霉毒素B1和玉米赤霉烯酮的新型霉菌毒素纯化系统

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摘要

In this study, we developed a novel tool for purifying two mycotoxins, aflatoxin B1 (AFB1) and zearalenone (ZEN), in feed. This system utilized monoclonal antibodies (mAbs) against AFB1 and ZEN, and magnetic nanoparticles (MNPs). Among ten MNPs with different diameters and functional groups, a 100-nm diameter MNP (fMA) conjugated to an amine group (-NH2) was found to be optimum for coupling with mAbs. The optimal mAb concentrations for coupling to the fMA along with mycotoxin purification capacities of the fMA-mAb conjugates (fMA-AFB1 and fMA-ZEN) were determined. A comparison of mean recovery rates (from corn and product X feed) between the fMA-mAb conjugates and immunoaffinity columns (IAC-AFB1 and IAC-ZEN) showed that the rate for fMA-AFB1 (90~92% and 81~88%) was higher (p > 0.05) than that of IAC-AFB1 (81~84% and 72~78%) for AFB1 (5, 10, 15 ng/mL), and the rate for fMA-ZEN (99~100% and 92~94%) was significantly higher (p < 0.01) than that of IAC-ZEN (86~88% and 81~88%) for ZEN (10, 25, 50 ng/mL) except at a concentration of 10 ng/mL, demonstrating the remarkable purification efficiency of the novel fMA-mAb method. Additionally, mycotoxin purification was much faster using our novel method (approx. 5 min) than the IAC-based technique (> 30 min). This study suggests that the novel purification system we developed would be a useful tool for monitoring and regulating mycotoxin contamination in feed, and replace IAC methods.
机译:在这项研究中,我们开发了一种纯化饲料中两种霉菌毒素黄曲霉毒素B1(AFB1)和玉米赤霉烯酮(ZEN)的新颖工具。该系统利用了针对AFB1和ZEN的单克隆抗体(mAb),以及磁性纳米颗粒(MNP)。在十个具有不同直径和官能团的MNP中,发现与胺基(-NH2)共轭的100 nm直径MNP(fMA)最适合与mAb偶联。确定了与fMA偶联的最佳mAb浓度以及fMA-mAb共轭物(fMA-AFB1和fMA-ZEN)的霉菌毒素纯化能力。比较fMA-mAb结合物和免疫亲和柱(IAC-AFB1和IAC-ZEN)的平均回收率(来自玉米和X饲料),表明fMA-AFB1的回收率(90〜92%和81〜88% )(5,10,15 ng / mL)高于IAC-AFB1(81〜84%和72〜78%),fMA-ZEN的比率(99〜100%)高于(IAC-AFB1)(81〜84%和72〜78%)和ZEN(10、25、50 ng / mL)的IAC-ZEN(86〜88%和81〜88%)的浓度(10%,25、50 ng / mL)明显高于IAC-ZEN(86〜88%和81〜88%)(p <0.01) / mL,证明了新型fMA-mAb方法的卓越纯化效率。此外,使用霉菌毒素的新方法(约5分钟)比基于IAC的技术(> 30分钟)纯化真菌毒素要快得多。这项研究表明,我们开发的新型纯化系统将成为监测和调节饲料中霉菌毒素污染的有用工具,并取代IAC方法。

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