Food Allergy Research and Resource Program, University of Nebraska, Lincoln, NE 68583-0919, USA.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2012;29(9):1347-55. doi: 10.1080/19440049.2012.698399. Epub 2012 Jul 24.
The major allergen parvalbumin was purified from cod muscle tissues, and polyclonal antibodies were raised towards it. The antibodies were tested for specificity and an enzyme-linked immunosorbent assay (ELISA) was developed using these antibodies. The ELISA was applied to measure parvalbumin in cod skin, the starting material for fish gelatin made from deep sea, wild fish. The ELISA was sufficiently sensitive (LLOQ = 0.8 ng ml(-1) in extracts, corresponding to 0.02 µg of parvalbumin per g of tissue), and did not cross-react with common food constituents. Fish gelatin, wine and beer, matrices for the potential use of this ELISA, did not cause disturbance of the assay performance. The data show that the parvalbumin content in cod muscle tissue is 6.25 mg g(-1), while the skins contained considerably less, 0.4 mg g(-1). Washing of the skins, a common industrial procedure during the manufacturing of fish gelatin, reduced the level of parvalbumin about 1000-fold to 0.5 µg g(-1), or 0.5 ppm. From 95 commercial lots of fish gelatin it is shown that 73 are below 0.02 µg g(-1) parvalbumin. From the other 22 lots, the one with the highest concentration contained 0.15 µg g(-1) of parvalbumin. These levels are generally assumed to be safe for fish-allergic individuals.
主要过敏原副肌球蛋白从鳕鱼肌肉组织中纯化，并针对其产生了多克隆抗体。测试了这些抗体的特异性，并使用这些抗体开发了酶联免疫吸附测定 (ELISA)。将 ELISA 应用于测量来自深海野生鱼的鱼明胶起始原料鳕鱼皮中的副肌球蛋白。该 ELISA 具有足够的灵敏度（LLOQ=0.8ng ml(-1) 在提取物中，相当于组织中每克 0.02 µg 的副肌球蛋白），并且不会与常见的食物成分发生交叉反应。鱼明胶、葡萄酒和啤酒，都是潜在应用此 ELISA 的基质，它们不会干扰分析性能。数据表明，鳕鱼肌肉组织中的副肌球蛋白含量为 6.25mg g(-1)，而鱼皮中的含量要少得多，为 0.4mg g(-1)。在鱼明胶生产过程中，通常进行的鱼皮清洗工序会使副肌球蛋白水平降低约 1000 倍，至 0.5 µg g(-1)，或 0.5ppm。从 95 批商业鱼明胶中可以看出，有 73 批的副肌球蛋白含量低于 0.02 µg g(-1)。在其他 22 批中，含量最高的一批含有 0.15 µg g(-1)的副肌球蛋白。这些水平通常被认为对鱼类过敏个体是安全的。
