Chassaigne Hubert, Nørgaard Jørgen V, Hengel Arjon J van
Food Safety and Quality Unit, Institute for Reference Materials and Measurements, European Commission - DG Joint Research Centre, Retieseweg 111, B-2440 Geel, Belgium.
J Agric Food Chem. 2007 May 30;55(11):4461-73. doi: 10.1021/jf063630e. Epub 2007 May 3.
An MS-based method, combining reversed-phase capillary liquid chromatography (capillary LC) with quadrupole time-of-flight tandem mass spectrometry (nano-ESI Q-TOF MS/MS), was developed with the aim of identifying a set of peptides that can function as markers for peanut allergens. Emphasis was given to the identification of the three major peanut allergens Ara h 1, Ara h 2, and Ara h 3, because these proteins are considered to represent >30% of the total protein content of peanut and are directly relevant for the allergenic potential of this food. The analytical data obtained were used to perform databank searching in combination with de novo sequencing and led to the identification of a multitude of sequence tags for all three peanut allergens. Food processing such as roasting of peanuts is known to affect the stability of proteins and was shown to influence the detection of allergen sequence tags. The analysis of raw and roasted peanuts allowed the identification of five peanut-specific sequence tags that can function as markers of the specific allergenic proteins. For Ara h 1, two peptide markers were proposed, namely, VLEENAGGEQEER (m/z 786.88, charge 2+) and DLAFPGSGEQVEK (m/z 688.85, charge 2+), whereas for Ara h 2 only one peptide, RQQWELQGDR (m/z 439.23, charge 3+), was found to satisfy the required conditions. For Ara h 3, the two specific peptides, SPDIYNPQAGSLK (m/z 695.35, charge 2+) and SQSENFEYVAFK (m/z 724.84, charge 2+), were selected. Other peptides have been proposed as indicative for food processing.
开发了一种基于质谱的方法，将反相毛细管液相色谱（毛细管液相色谱）与四极杆飞行时间串联质谱（纳升电喷雾四极杆飞行时间串联质谱）相结合，旨在鉴定一组可作为花生过敏原标志物的肽段。重点是鉴定三种主要的花生过敏原Ara h 1、Ara h 2和Ara h 3，因为这些蛋白质被认为占花生总蛋白质含量的30%以上，并且与这种食物的致敏潜力直接相关。所获得的分析数据用于结合从头测序进行数据库搜索，并导致鉴定出所有三种花生过敏原的大量序列标签。已知诸如花生烘烤等食品加工过程会影响蛋白质的稳定性，并已证明会影响过敏原序列标签的检测。对生花生和烤花生的分析使得能够鉴定出五个可作为特定致敏蛋白标志物的花生特异性序列标签。对于Ara h 1，提出了两个肽段标志物，即VLEENAGGEQEER（m/z 786.88，电荷2+）和DLAFPGSGEQVEK（m/z 688.85，电荷2+），而对于Ara h 2，仅发现一个肽段RQQWELQGDR（m/z 439.23，电荷3+）满足所需条件。对于Ara h 3，选择了两个特定的肽段，SPDIYNPQAGSLK（m/z 695.35，电荷2+）和SQSENFEYVAFK（m/z 724.84，电荷2+）。还提出了其他肽段作为食品加工的指示物。
