Dietrich Richard, Märtlbauer Erwin
Department of Veterinary Sciences, Ludwig Maximilians-Universität München, Schönleutnerstraße 8, 85764, Oberschleißheim, Germany.
Mycotoxin Res. 2015 Nov;31(4):185-90. doi: 10.1007/s12550-015-0229-3. Epub 2015 Sep 17.
Mycophenolic acid (MPA) is frequently found, often in high concentrations, in a broad range of food and feed matrices. Apart from the well-known contamination of blue-veined cheeses caused by the use of toxinogenic Penicillium roqueforti strains for manufacturing, a broad range of other Penicillium spp. is able to produce this immunosuppressive toxin. Therefore, MPA has been proposed to be a suitable marker for Penicillium-infected food commodities. In the present work, a high-affinity monoclonal antibody (mAb) for the specific detection of MPA was developed by immunizing mice with a MPA-protein conjugate coupled by an activated ester method. Under the conditions of a direct competitive enzyme immunoassay (EIA), 50% inhibition and detection limits of MPA standard curves were 1.2 and 0.3 ng/ml, respectively. Furthermore, the mAb could be successfully employed for the production of an immunoaffinity (IA) column enabling the efficient enrichment of MPA from processed foodstuffs. By combining the IA clean-up with a polyclonal antibody-based EIA, an ultrasensitive analysis method could be established which allowed the reliable and reproducible detection of MPA in artificially contaminated tomato ketchup as a model matrix at concentrations as low as 0.1 ng/g.
麦考酚酸(MPA)在多种食品和饲料基质中经常被发现,且浓度往往较高。除了因使用产毒素的罗克福青霉菌株制造蓝纹奶酪而导致的众所周知的污染外,其他多种青霉菌也能够产生这种免疫抑制毒素。因此,MPA被提议作为受青霉菌感染的食品的合适标志物。在本研究中,通过用活化酯法偶联的MPA - 蛋白质偶联物免疫小鼠,开发了一种用于特异性检测MPA的高亲和力单克隆抗体(mAb)。在直接竞争酶免疫分析(EIA)条件下,MPA标准曲线的50%抑制率和检测限分别为1.2和0.3 ng/ml。此外,该单克隆抗体可成功用于制备免疫亲和(IA)柱,从而能够从加工食品中有效富集MPA。通过将IA净化与基于多克隆抗体的EIA相结合,可以建立一种超灵敏分析方法,该方法能够在人工污染的番茄酱(作为模型基质)中可靠且可重复地检测低至0.1 ng/g浓度的MPA。