鳜鱼致敏蛋白的识别及酶法水解的脱敏分析

Identification of IgE-binding protein from mandarin fish and antigenicity reduced by proteinases hydrolysis

试验采用聚丙烯酰胺凝胶电泳对鳜鱼蛋白提取液中不同分子质量的蛋白组分进行分离,在阳性血清池建立的基础上,运用转移电泳、免疫印迹以及免疫印迹抑制试验识别特异性吸附蛋白条带。进而采取不同来源的6种食品级蛋白酶对鳜鱼蛋白水解脱敏,将中性甲醛电位滴定法和间接酶联免疫吸附法测定的水解度和抗原降低率作为评判指标优选出最佳水解酶。最终得到分子质量为60、50、34 ku和17 ku的4条蛋白条带可以与IgE阳性结合,最佳水解酶为木瓜蛋白酶。

SDS-PAGE was employed to further identify the relative contents of major allergens of mandarin fJsh（SinJperca chuatsi） protein. On the basis of positive serum, transfer electrophoresis, western blotting and immunoinhibition tests were used to quantify specific adsorption of protein bands. And then six different sources of food-grade proteases were adopted to hydrolyze allergens in order to hyposensitization. As two indicators of the optimized hydrolase, degree of hydrolysis and lower rates of allergens were checked by Neutral formaldehyde titration and indirect enzyme-linked immunosorbent assay. Finally the molecular weight of 60 ku, 5 ku, 3 ku and 1 ku protein bands can be connected with the specific immunoglobulin E. Secondly the best enzyme was papain.