【摘要】 目的 探究南美白对虾虾壳蛋白稳定性及致敏性，鉴定致敏蛋白的种类并预测抗原表位。方法 盐析法提取虾壳粗蛋白后对其进行热处理、酸碱处理及模拟胃液消化处理，利用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行分析。利用血清学方法识别虾壳蛋白中与过敏患者血清特异性抗体结合的条带，利用液相色谱-串联质谱对结合的蛋白条带进行分析。运用生物信息学分析工具对质谱结果进行致敏性及B细胞线性表位预测。结果 虾壳蛋白热稳定性较好，但在酸碱环境中稳定性较差且不耐消化。虾壳蛋白中与过敏患者血清特异性抗体结合的蛋白的分子量在75 kDa左右。虾壳中主要存在的潜在致敏蛋白为血蓝蛋白和β-1,3-葡聚糖结合蛋白。结论 血蓝蛋白可能是存在于南美白对虾虾壳中的大分子量致敏蛋白，具备热稳定性但易被胃液消化，其抗原表位已通过生物信息学方法初步预测。本研究为虾壳产品的过敏性评价及低敏化加工技术提供了理论基础。更多还原

【Abstract】 Objective To explore the stability and allergenicity of Litopenaeus vannamei shell protein, identify the categories of allergenic proteins, and predict the antigen epitopes.Methods After extracting crude protein from shrimp shells using salt precipitation, L. vannamei shell protein was treated sequentially with heat, acid/base, and simulated gastric digestion, followed by electrophoretic analysis. Serological method was used to identify bands that L. vannamei shrimp shell protein bind specifically to antibodies from the serum of allergic patients, and then the bound protein bands were analyzed using liquid chromatography-tandem mass spectrometry. Bioinformatics analysis were employed for allergenicity analysis and B-cell linear epitope prediction, based on mass spectrometry results.Results L. vannamei shell protein was relatively stable to heat, however it was unstable in acidic and alkaline conditions and was easily digested by simulated gastric fluid. The molecular weights of the proteins in L. vannamei shell that specifically bound to antibodies from allergic patients were approximately 75 kDa. The main potential allergenic proteins present in L. vannamei shells were hemocyanin and β-1,3-glucan binding protein.Conclusion The existence of hemocyanin in the shells of L. vannamei, which may be an allergenic protein with high molecular weight and exhibited thermal stability but being easily digested by gastric juic. This study provides evidence for the allergenicity evaluation of L. vannamei shell products and the necessity for their low-allergen processing.更多还原