Fish is one of the most common foods that cause allergic reactions. The study of cross-reactivity among fishes using mass spectrometry (MS) is still limited. We developed a strategy using microfluidic chips coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to evaluate cross-reactivity among fishes. The protocol employed commercial magnetic beads functionalized with anti-human IgE antibodies to carry out the IgEs immunomagnetic separation in blood samples, followed by the capture of allergens from seafood protein extracts in a single-straight microfluidic channel. After elution, the captured allergens were digested and identified by MALDI-TOF MS and high-performance liquid chromatography-tandem mass spectrometry and validated by enzyme-linked immunosorbent assay (ELISA). An investigation of the reproducibility revealed that the protocol can sense well the allergens in a food matrix. Seven fish species were analyzed to evaluate the allergic cross-reactivity among fishes. The commercial ELISA test gave consistent results with the presently developed strategy when the same allergenicity test was performed. Parvalbumins were detected from five of the seven analyzed fishes. The sequence alignment of parvalbumins revealed that the similarity of parvalbumins identified from the analyzed fishes is larger than 64%. Boiling may reduce the allergenicity of fish, as demonstrated by a marginal diminish in the parvalbumin content of crucian carp (Carassius carassius) muscle when boiling with water. The method can potentially be used to predict allergic cross-reactivity among fish species, provide advice and guidance to individuals with a history of seafood allergy, and ensure food safety in the food allergy community.

中文翻译：

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微流控芯片和 MALDI-TOF MS 评估鱼类过敏性交叉反应

鱼是最常见的引起过敏反应的食物之一。使用质谱 (MS) 研究鱼类之间的交叉反应仍然有限。我们开发了一种策略，使用微流控芯片结合基质辅助激光解吸/电离飞行时间质谱 (MALDI-TOF MS) 来评估鱼类之间的交叉反应。该协议采用商业磁珠，用抗人 IgE 抗体功能化，在血液样本中进行 IgEs 免疫磁性分离，然后在单直微流体通道中从海鲜蛋白提取物中捕获过敏原。洗脱后，捕获的过敏原通过MALDI-TOF MS和高效液相色谱-串联质谱进行消化鉴定，并通过酶联免疫吸附试验（ELISA）进行验证。对可重复性的调查表明，该协议可以很好地感知食物基质中的过敏原。分析了七种鱼类以评估鱼类之间的过敏交叉反应。当进行相同的过敏性测试时，商业 ELISA 测试给出了与目前开发的策略一致的结果。从所分析的七种鱼类中的五种中检测到小白蛋白。小白蛋白的序列比对显示，从分析的鱼类中鉴定出的小白蛋白的相似性大于 64%。煮沸可以降低鱼的致敏性，鲫鱼的小白蛋白含量略有减少就证明了这一点。当进行相同的过敏性测试时，商业 ELISA 测试给出了与目前开发的策略一致的结果。从所分析的七种鱼类中的五种中检测到小白蛋白。小白蛋白的序列比对显示，从分析的鱼类中鉴定出的小白蛋白的相似性大于 64%。煮沸可以降低鱼的致敏性，鲫鱼的小白蛋白含量略有减少就证明了这一点。当进行相同的过敏性测试时，商业 ELISA 测试给出了与目前开发的策略一致的结果。从所分析的七种鱼类中的五种中检测到小白蛋白。小白蛋白的序列比对显示，从分析的鱼类中鉴定出的小白蛋白的相似性大于 64%。煮沸可以降低鱼的致敏性，鲫鱼的小白蛋白含量略有减少就证明了这一点。Carassius carassius ) 与水一起煮沸时的肌肉。该方法可用于预测鱼类之间的过敏交叉反应，为有海鲜过敏史的个体提供建议和指导，并确保食物过敏社区的食品安全。