Masiri Jongkit, Benoit Lora, Katepalli Madhu, Meshgi Mahzad, Cox David, Nadala Cesar, Sung Shao-Lei, Samadpour Mansour
Molecular Epidemiology, Inc. (MEI) , 15300 Bothell Way N.E., Lake Forest Park, Washington 98155, United States.
IEH Laboratories and Consulting Group, Inc. (IEH) , 15300 Bothell Way N.E., Lake Forest Park, Washington 98155, United States.
J Agric Food Chem. 2016 May 11;64(18):3678-87. doi: 10.1021/acs.jafc.5b06085. Epub 2016 Apr 27.
Gluten derived from wheat and related Triticeae can induce gluten sensitivity as well as celiac disease. Consequently, gluten content in foods labeled "gluten-free" is regulated. Determination of potential contamination in such foods is achieved using immunoassays based on monoclonal antibodies (mAbs) that recognize specific epitopes present in gluten. However, food-processing measures can affect epitope recognition. In particular, preparation of wheat protein isolate through deamidation of glutamine residues significantly limits the ability of commercial gluten testing kits in their ability to recognize gluten. Adding to this concern, evidence suggests that deamidated gluten imparts more pathogenic potential in celiac disease than native gluten. To address the heightened need for antibody-based tools that can recognize deamidated gluten, we have generated a novel mAb, 2B9, and subsequently developed it as a rapid lateral flow immunoassay. Herein, we report the ability of the 2B9-based lateral flow device (LFD) to detect gluten from wheat, barley, and rye and deamidated gluten down to 2 ppm in food as well as its performance in food testing.
源自小麦和相关小麦族的麸质可引发麸质敏感性以及乳糜泻。因此,标有“无麸质”的食品中的麸质含量受到监管。使用基于单克隆抗体(mAb)的免疫测定法来检测此类食品中的潜在污染物,这些单克隆抗体可识别麸质中存在的特定表位。然而,食品加工措施会影响表位识别。特别是,通过谷氨酰胺残基脱酰胺作用制备小麦蛋白分离物会显著限制商业麸质检测试剂盒识别麸质的能力。此外,有证据表明,脱酰胺麸质在乳糜泻中比天然麸质具有更大的致病潜力。为满足对能够识别脱酰胺麸质的基于抗体的工具的更高需求,我们制备了一种新型单克隆抗体2B9,并随后将其开发成一种快速侧向流动免疫测定法。在此,我们报告基于2B9的侧向流动装置(LFD)检测食品中小麦、大麦和黑麦麸质以及脱酰胺麸质低至2 ppm的能力及其在食品检测中的性能。