Kim Hyun-Jung, Kim Sung-Hee, Lee Jin-Kyu, Choi Cheong-Up, Lee Hee-Soo, Kang Hwan-Goo, Cha Sang-Ho
Animal, Plant and Fisheries Quarantine and Inspection Agency, Anyang 480-757, Korea.
J Vet Sci. 2012 Dec;13(4):363-9. doi: 10.4142/jvs.2012.13.4.363.
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 (-NH(2)) 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 (9092% and 8188%) was higher (p > 0.05) than that of IAC-AFB1 (8184% and 7278%) for AFB1 (5, 10, 15 ng/mL), and the rate for fMA-ZEN (99100% and 9294%) was significantly higher (p < 0.01) than that of IAC-ZEN (8688% and 8188%) 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.
在本研究中,我们开发了一种用于净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)的新型工具。该系统利用了针对AFB1和ZEN的单克隆抗体(mAbs)以及磁性纳米颗粒(MNPs)。在十种具有不同直径和官能团的MNPs中,发现一种与胺基(-NH(2))偶联的直径为100纳米的MNP(fMA)最适合与mAbs偶联。确定了与fMA偶联的最佳mAb浓度以及fMA - mAb偶联物(fMA - AFB1和fMA - ZEN)的霉菌毒素净化能力。fMA - mAb偶联物与免疫亲和柱(IAC - AFB1和IAC - ZEN)之间的平均回收率(来自玉米和产品X饲料)比较表明,对于AFB1(5、10、15纳克/毫升),fMA - AFB1的回收率(9092%和8188%)高于IAC - AFB1(8184%和7278%)(p > 0.05);对于ZEN(10、25、50纳克/毫升),除了10纳克/毫升的浓度外,fMA - ZEN的回收率(99100%和9294%)显著高于IAC - ZEN(8688%和8188%)(p < 0.01),这表明新型fMA - mAb方法具有显著的净化效率。此外,使用我们的新方法(约5分钟)进行霉菌毒素净化比基于IAC的技术(>30分钟)要快得多。本研究表明,我们开发的新型净化系统将是监测和调控饲料中霉菌毒素污染的有用工具,并可取代IAC方法。