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目的预测金黄色葡萄球球菌Newman株IsdA蛋白B细胞线性抗原表位,并对所预测B细胞表位的免疫反应性进行初步鉴定。方法以金黄色葡萄球球菌Newman株IsdA蛋白为对象,采用NCBI提供的蛋白质查询程序确定IsdA蛋白的氨基酸序列,然后使用网络在线预测软件SOPMA分析IsdA蛋白质二级结构;使用IEDB网络服务器预测IsdA蛋白B细胞线性表位和Th细胞表位;使用VectorNTI10.0软件分析IsdA蛋白序列中易形成B细胞表位的氨基酸-缬氨酸、亮氨酸及半胱氨酸所在的位置。综合以上分析结果获得所预测B细胞线性表位。以预测表位序列为参考,人工合成表位多肽,通过肽间接ELISA法验证所预测表位与IsdA多克隆抗体血清的免疫反应性。结果 IsdA存在潜在的B细胞线性抗原表位,所预测表位区分别位于N末端P_(120-140)、P_(181-200)、P_(210-230)和P_(300-320),间接ELISA检测rIsdA多克隆抗体血清中存在着可与P_(210-230)、P_(300-320)结合的组分。结论 IsdA蛋白潜在的B细胞线性表位位于N末端P_(210-230)和P_(300-320)区域,具有与特异性血清结合的能力,为金黄色葡萄球菌的表位疫苗研发奠定了基础。
Objective To predict the linear epitope of IsdA protein B of Staphylococcus aureus Newman strain and to identify the immunoreactivity of predicted B cell epitopes. Methods The IsdA protein of Staphylococcus aureus Newman strain was used to determine the amino acid sequence of IsdA protein by using the protein query program provided by NCBI. Then the secondary structure of IsdA protein was analyzed by online prediction software SOPMA. The IEDB network server was used to predict IsdA protein B Linear epitopes of cells and Th cell epitopes; the amino acid positions of valine, leucine and cysteine that are easy to form B cell epitope in IsdA protein sequence were analyzed by VectorNTI10.0 software. Based on the above analysis results, the predicted linear B-cell epitopes were obtained. Using the predicted epitope sequence as a reference, the epitope peptide was synthesized by artificial ELISA and the immunoreactivity of the predicted epitope to IsdA polyclonal antibody serum was verified by the peptide indirect ELISA. Results There was a potential linear epitope of B cells in IsdA. The predicted epitopes were located at the N-terminal P_ (120-140), P_ (181-200), P_ (210-230) and P_ (300-320) The serum of rIsdA polyclonal antibody detected by ELISA showed that there was a component that could bind to P 210-230 and P 300-320. Conclusion The potential B cell linear epitopes of IsdA protein are located in the N-terminal P 210-230 and P 300-320 regions, which have the ability to bind with specific serum and lay a foundation for the development of the epitope vaccine of S. aureus .