榛子主要过敏原Cor h 1单克隆抗体识别抗原表位区域的确定

目的 确定已获得的四株榛子主要过敏原Cor h 1单克隆抗体识别的抗原表位区域。方法 构建谷胱甘肽疏基转移酶 (GST)与Cor h 1不同截短体的表达载体并表达GST-Cor h 1融合蛋白, 采用ELISA和Western blot检测单抗与不同融合蛋白的反应, 并结合DNAstar表位分析软件推知不同单抗的抗原识别表位区域。结果 确定了四株Cor h 1单抗的抗原识别表位区域, 分别是4G7和2H3抗体的抗原表位位于Cor h 1第120~126位氨基酸, 5F2和1G4抗体的抗原表位位于Cor h 1第43~52位氨基酸。结论 四株Cor h 1单抗的抗原识别表位区域的确定, 为Cor h 1的进一步研究和食品安全检测奠定了基础。

Identification of hazelnut antigenic epitope regions recognized by anti-Cor h 1 monoclonal antibodies

Objective To identify the antigenic epitope regions recognized by anti-Cor h 1 monoclonal antibodies. Methods Five expression vectors of truncated Cor h 1 were constructed and the truncated Cor h 1-GST fusion proteins were expressed in E. coli. The binding activities of anti-Cor h 1 monoclonal antibodies (mAbs) with truncated Cor h 1 were analyzed by ELISA and Western blot. According to the speculative epitope regions, expression vectors of potential epitopes were constructed and the GST-epitope fusion proteins were expressed in E. coli. The fusion proteins were analyzed by ELISA and Western blot with mAbs to confirm the epitope regions, with the DNAstar epitope analysis software. Results Epitope regions recognized by four anti-Cor h 1 mAbs were identified. The epitopes recognized by 4G7 and 2H3 were located in the region from 120th to 126th amino acids of Cor h 1, 5F2 and 1G4 in the region from 43th to 54th amino acids. Conclusion The identification of epitope regions recognized by four anti-Cor h 1 mAbs will benefit the research on Cor h 1 and the food safe examination.