College of Food Science and Engineering, Shanghai Ocean University, Hucheng Ring Road 999, Pudong New District, 201306 Shanghai, China.
Analyst. 2022 Sep 12;147(18):4063-4072. doi: 10.1039/d2an00813k.
The main objective of the current study is to establish an on-chip strategy to analyze the allergens existing in the myofibrillar proteins of seafood matrices. On-chip immunomagnetic separation (IMS) of allergens derived from large yellow croaker (), swimming crab () and black tiger shrimp (), as model seafoods, was studied. The analysis of the tryptic digests of on-chip captured proteins, combined with protein database searches, was performed for the identification of allergens by MALDI-TOF MS with a 0% false-positive rate. By using the sera of ten seafood-allergic patients, six allergens were identified from the myofibrillar proteins of the large yellow croaker. The present protocol was successfully validated by ELISA and can be applied in blood tests for food allergies with a single drop of blood. The current protocol is presumed to be applied in the identification of potential allergens in other organisms.
本研究的主要目的是建立一种用于分析海鲜基质中肌球蛋白过敏原的芯片策略。对大黄鱼()、梭子蟹()和黑虎虾()三种模式海鲜来源的过敏原进行了芯片免疫磁分离(IMS)研究。采用 MALDI-TOF MS 结合蛋白酶解产物数据库检索,对芯片捕获蛋白进行分析,实现了过敏原的鉴定,其假阳性率为 0%。利用 10 例海鲜过敏患者的血清,从大黄鱼肌球蛋白中鉴定出 6 种过敏原。通过 ELISA 对该方法进行验证,该方法仅需一滴血即可用于食物过敏的血液检测。该方案有望应用于其他生物中潜在过敏原的鉴定。