Quantitation of Soybean Allergens Using Tandem Mass Spectrometry
Quantitation of Soybean Allergens Using Tandem Mass Spectrometry
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DOI：
10.1021/pr100913w
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发表时间：
2011-02-01
影响因子：
4.4
通讯作者：
Thelen, Jay J.
中科院分区：
文献类型：
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作者：
Houston, Norma L.;Lee, Dong-Gi;Thelen, Jay J.
大豆（Glycine max）种子含有一些对人类和动物过敏的蛋白质。然而，这些过敏原的浓度及其在种质之间的表达变异性目前尚不清楚。为了解决这个问题，10过敏原定量从20个非转基因商业大豆品种使用平行的，无标记的质谱方法。通过光谱计数进行相对定量，并使用多反应监测（MRM）进行绝对定量，合成的同位素标记的肽作为内标。在相对定量分析过程中，鉴定了10种目标过敏原，其中5种过敏原的表达水平高于牛血清白蛋白（BSA）内标物的技术变异（类似于11%），表明品种间存在表达差异。为了证实这一观察结果，使用MRM对每个品种的这些过敏原进行绝对定量。10种过敏原中的8种被定量测定其在种子中的浓度，范围约为0.5至5.7 μ g/mg大豆蛋白。MRM分析将BSA内标的技术方差降低至约7%，并证实了20个品种中4种过敏原的差异表达。这是对所有主要大豆过敏原的首次定量评估。结果表明，20个大豆品种的过敏原总量基本相似。
Soybean (Glycine max) seed contain some proteins that are allergenic to humans and animals. However, the concentration of these allergens and their expression variability among germplasms is presently unknown. To address this problem, 10 allergens were quantified from 20 nongenetically modified commercial soybean varieties using parallel, label-free mass spectrometry approaches. Relative quantitation was performed by spectral counting and absolute quantitation was performed using multiple reaction monitoring (MRM) with synthetic, isotope-labeled peptides as internal standards. During relative quantitation analysis, 10 target allergens were identified, and five of these allergens showed expression levels higher than technical variation observed for bovine serum albumin (BSA) internal standard (similar to 11%), suggesting expression differences among the varieties. To confirm this observation, absolute quantitation of these allergens from each variety was performed using MRM. Eight of the 10 allergens were quantified for their concentration in seed and ranged from approximately 0.5 to 5.7 mu g/mg of soy protein. MRM analysis reduced technical variance of BSA internal standards to approximately 7%, and confirmed differential expression for four allergens across the 20 varieties. This is the first quantitative assessment of all major soybean allergens. The results show the total quantity of allergens measured among the 20 soy varieties was mostly similar.