pH值和酶作用对鲩鱼过敏原免疫学活性的影响研究

Influence of pH and enzymolysis on the immunogenicity of Ctenopharyngodon idellus allergens

目的研究pH值、胃蛋白酶和胰蛋白酶作用对鲩鱼过敏原免疫学活性的影响。方法采用不同Coca's液提取制备鲩鱼总蛋白,以鲩鱼过敏患者阳性血清为一抗用Western Blotting方法分析pH值、胃蛋白酶和胰蛋白酶以及多因素多重作用对鲩鱼过敏原免疫学特性的影响。结果HCl溶液、胃蛋白酶溶液、胰蛋白酶溶液以及多种因素的多重作用对鲩鱼蛋白均有不同程度的降解作用,且均可以显著降低鲩鱼过敏原的免疫学活性,但一些蛋白具有很高的稳定性,如M_r为35 000和23 000的蛋白。受制备过程的影响,鲩鱼蛋白中出现新的阳性过敏原条带,如M_r为27 000和20 000的阳性过敏原条带(pH 1.0 HCl溶液)。结论鲩鱼含有稳定性较高的潜在过敏原,该类潜在过敏原对热、酸、酶解等作用均有很强的耐受性。

This study aimed to evaluate the influence of pH and pepsin/trypsin enzymolysis on the immunogenicity of Ctenopharyngodon idellus allergens.Total proteins from Ctenopharyngodon idellus were extracted by different Coca＇s solutions,separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE）,and detected with Coomassie Brillant Blue staining. The immunogenicity of allergens treated with acid,pepsin or/and trypsin was analyzed by Western blotting using Ctenopharyngodon idellus allergic patients＇ sera.Components of the total proteins extracted from fishes treated with HCl solution,pepsin solutions or and trypsin solutions were less than that from raw fish and were different from each other.Western blotting analysis showed pH,pepsin, trypsin and heating had remarkable effects on the composition and immunogenicity of Ctenopharyngodon idellus allergens.But some pre-allergens in Ctenopharyngodon idellus still showed stable immunogenicity.All these results mean that there are some stable proteins in Ctenopharyngodon idellus,which are tolerant to heating,acidity,enzymolysis,even all of them.