College of Food Science and Engineering, Shanghai Ocean University, Hucheng Ring Road 999, Pudong New District, 201306 Shanghai, China.
Department of Chemistry, Fudan University, Songhu Road 2005, Yangpu District, 200433 Shanghai, China.
J Agric Food Chem. 2021 Nov 3;69(43):12909-12918. doi: 10.1021/acs.jafc.1c04723. Epub 2021 Oct 6.
We developed a strategy using immunomagnetic separation (IMS) coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to test seafood allergens. The protocol employed commercial magnetic beads (MBs) functionalized with anti-human IgE antibodies to carry out the IMS of IgEs in blood samples, followed by capture of allergens from seafood protein extracts for allergy analysis. After elution, the captured allergens were identified by MALDI-TOF MS and HPLC-MS/MS. The non-specific adsorption of MBs to biomolecules, the reproducibility and sensitivity of the protocol were investigated. The method shows consistent results with enzyme-linked immunosorbent assay tests. The false positive rate of the present method for the allergy test is 0%. The protocol was applied to detect the allergens in greasy-back shrimp for checking the allergenicity of patients' serum. Cooking fish as soup may effectively decrease the allergenicity. The method can be potentially used to identify unknown allergens of seafood to ensure the safety of allergic patients.
我们开发了一种使用免疫磁珠分离（IMS）结合基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）的策略来检测海鲜过敏原。该方案采用了商业化的磁珠（MBs），其表面用抗人 IgE 抗体修饰，用于在血液样本中进行 IgE 的 IMS，然后从海鲜蛋白提取物中捕获过敏原进行过敏分析。洗脱后，通过 MALDI-TOF MS 和 HPLC-MS/MS 鉴定捕获的过敏原。研究了 MBs 对生物分子的非特异性吸附、该方案的重现性和灵敏度。该方法与酶联免疫吸附试验测试结果一致。本方法用于过敏测试的假阳性率为 0%。该方案用于检测油腻虾中的过敏原，以检查患者血清的致敏性。将鱼煮成汤可能会有效降低其致敏性。该方法可用于鉴定海鲜的未知过敏原，以确保过敏患者的安全。
