食品科学 ›› 2018, Vol. 39 ›› Issue (22): 256-262.doi: 10.7506/spkx1002-6630-201822039

• 安全检测 • 上一篇    下一篇

生物膜干涉法检测花生蛋白与花生过敏患者血清IgE的结合能力

张英1,2,李坤1,3,颜琪1,2,陈红兵1,4,吴志华1,4,*   

  1. (1.南昌大学?食品科学与技术国家重点实验室,江西?南昌 330047;2.南昌大学食品学院,江西?南昌 330031;3.南昌大学资源环境与化工学院,江西?南昌 330031;4.南昌大学中德联合研究院,江西?南昌 330047)
  • 出版日期:2018-11-25 发布日期:2018-11-21
  • 基金资助:
    国家高技术研究发展计划(863计划)项目(2013AA102205);国家自然科学基金面上项目(31771924); 国家自然科学基金地区科学基金项目(31660446); 江西省主要学科学术和技术带头人资助计划项目(20172BCB22003); 食品科学与技术国家重点实验室目标导向课题(SKLF-ZZA-201612); 食品科学与技术国家重点实验室自主探索项目(SKLF-ZZB-201514)

Detection of the Binding Capacity between Peanut Protein and Serum IgE from Peanut Allergy Sufferers Using Biolayer Interferometry

ZHANG Ying1,2, LI Kun1,3, YAN Qi1,2, CHEN Hongbing1,4, WU Zhihua1,4,*   

  1. (1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; 2. School of Food Science and Technology, Nanchang University, Nanchang 330031, China; 3. School of Environment & Chemical Engineering, Nanchang University, Nanchang 330031, China; 4. Sino-German Joint Research Institute, Nanchang University, Nanchang 330047, China)
  • Online:2018-11-25 Published:2018-11-21

摘要: 探索基于生物膜干涉技术对花生蛋白与花生过敏患者血清中免疫球蛋白E(immunoglobulin E,IgE)的结合能力进行检测的方法。利用链霉亲和素(streptavidin,SA)标记的传感器、生物素化的羊抗人IgE抗体、花生过敏患者血清池以及花生蛋白建立了一种测定花生蛋白与花生过敏患者血清IgE结合能力的新方法,优化检测条件为抗体1∶100稀释后线下固化20?min,血清1∶10稀释后过夜结合,完成传感器修饰。在线洗基线后用质量浓度为1?mg/mL的花生蛋白与传感器结合3?600?s,解离120?s。利用该法对不同热加工后花生蛋白与患者血清IgE的结合能力进行评估,并与常用的酶联免疫吸附实验(enzyme linked immunosorbent assay,ELISA)检测进行比较。结果表明,本方法可以直接评估过敏原蛋白与血清IgE的结合能力,与ELISA结果相关系数达到0.91。热加工中,油炸处理提高了花生蛋白的IgE结合能力,水煮和烘烤降低了花生蛋白的IgE结合能力,且去壳热加工比带壳热加工花生的蛋白的IgE结合能力更强。

关键词: 花生蛋白, IgE, 生物膜干涉技术, IgE结合能力, 热加工

Abstract: In the present study, an approach to detect the binding capacity between peanut protein and serum IgE from patients with peanut allergy was explored using biolayer interferometry (BLI). For this purpose, the experiment was designed using a streptavidin (SA) dip and read biosensor, biotin labeled goat anti-human IgE antibody, the serum pool of patients with peanut allergy and peanut proteins. The optimized conditions for sensor modification were as follows: immobilization of the 100-fold diluted IgE for 20 min before being bound to the 10-fold diluted serum overnight. After being washed to baseline online, peanut protein (1 mg/mL) was bound to the sensor for 3 600 s and then dissociated for 120 s. This method was applied to detect the IgE binding capacity of peanut protein subjected to different thermal treatments and compare with enzyme linked immunosorbent assay (ELISA). The results showed that this method could directly evaluate the IgE binding capacity of allergenic protein and had good consistency with the ELISA results with correlation coefficient of 0.91. After frying treatment, the IgE binding capacity of peanut protein increased but decreased after boiling and roasting treatments. Protein extracted from thermally processed peanuts without shells had a stronger IgE binding capacity than that from thermally processed peanuts with shells.

Key words: peanut protein, IgE, biolayer interferometry, IgE binding capacity, thermal processing

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