Peanut is a major cause of severe IgE-mediated food allergic reactions, which can be exacerbated by factors, such as exercise, that may increase allergen uptake into the circulation. Enzyme-linked immunosorbent assays (ELISAs) have been used to determine allergen uptake into serum, but there are concerns over their specificity and a confirmatory method is required. Mass spectrometry (MS) methods have the potential to provide rigorous alternatives for allergen determination. A suite of peptide targets representing the major clinically relevant peanut allergens previously applied in food analysis were used to develop a targeted multiple reaction monitoring (MRM) method for determination of peanut in serum. Depletion of serum using affinity chromatography was found to be essential to allow detection of the peptide targets. A comparison of triple quadrupole and Q-TOF methods showed that one Ara h 2 peptide was only detected by the Q-TOF, the other peptide targets giving similar assay sensitivities with both MS platforms, although transitions for all the peptides were detected more consistently with the Q-TOF. The Q-TOF MRM assay detected peanut from spiked serum more effectively than the triple quadrupole assay, with Ara h 3 being detected down to 3 mg total peanut protein/L of serum, comparable with an Ara h 3-specific ELISA. The poor recoveries observed for both methods are likely due to loss of peanut immune complexes during the serum depletion process. Nevertheless, the Q-TOF MRM method has much promise to confirm the uptake of peanut proteins in serum samples providing immune complexes can be disrupted effectively prior to depletion. Graphical abstract.

中文翻译：

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使用四极杆和飞行时间质谱法测定血清中花生（花生）过敏原的多反应监测方法。

花生是严重的IgE介导的食物过敏反应的主要原因，运动等因素可能会加剧该过敏反应，例如运动可能会增加过敏原对循环系统的吸收。酶联免疫吸附测定（ELISA）已用于确定血清中的过敏原摄取，但是对它们的特异性存在担忧，需要一种验证方法。质谱（MS）方法有可能为确定过敏原提供严格的替代方法。代表先前在食品分析中应用的主要临床相关花生过敏原的一系列肽靶标可用于开发目标多反应监测（MRM）方法，用于测定血清中的花生。发现使用亲和色谱法消耗血清对于检测肽靶标至关重要。对三重四极杆和Q-TOF方法的比较显示，仅通过Q-TOF可检测到一种Ara h 2肽，而另一种肽靶标在两种MS平台上均具有相似的检测灵敏度，尽管在使用Q-TOF。Q-TOF MRM分析比三重四极杆检测更有效地从加标血清中检测出花生，检测到的Ara h 3含量低至3 mg花生总蛋白/ L，与Ara h 3特异性ELISA相当。两种方法均观察到较差的回收率，这可能是由于血清消耗过程中花生免疫复合物的损失。然而，如果可以在消耗前有效地破坏免疫复合物，那么Q-TOF MRM方法有望证实血清样品中花生蛋白的摄取。图形概要。