College of Food Science and Engineering, Shanghai Ocean University, Hucheng Ring Road 999, Pudong New District, Shanghai 201306, China.
J Agric Food Chem. 2022 Jun 22;70(24):7525-7534. doi: 10.1021/acs.jafc.2c02433. Epub 2022 Jun 13.
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 () 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.
鱼类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。我们开发了一种使用微流控芯片与基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）相结合的策略来评估鱼类之间的交叉反应。该方案采用商业磁性珠，其功能化有抗人 IgE 抗体，用于在血液样本中进行 IgE 免疫磁分离，然后在单个直微流控通道中捕获海鲜蛋白提取物中的过敏原。洗脱后，用 MALDI-TOF MS 和高效液相色谱-串联质谱对捕获的过敏原进行消化和鉴定，并通过酶联免疫吸附试验（ELISA）进行验证。对重现性的研究表明，该方案可以很好地检测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。当进行相同的变应原性测试时，商业 ELISA 测试与目前开发的策略得出一致的结果。从七种分析的鱼类中检测到副肌球蛋白。副肌球蛋白的序列比对表明，从分析鱼类中鉴定出的副肌球蛋白的相似性大于 64％。煮沸可能会降低鱼类的致敏性，因为在用水煮沸鲫鱼（）肌肉时，副肌球蛋白的含量略有减少。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。
