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目的筛选SARS病毒S蛋白的B细胞表位。方法使用SARS-CoV S DNA疫苗免疫BALB/c小鼠,获得SARS病毒S蛋白的免疫血清。人工合成包含169条部分氨基酸序列重叠的SARS-CoV S蛋白的多肽库。将多肽片段包被ELISA板,利用免疫小鼠血清,通过抗体结合试验来筛选SARS病毒S蛋白的线性B细胞表位。并将筛选结果与使用B细胞表位分析软件预测的结果进行比较。结果使用重叠肽合成法筛选到SARS-CoV蛋白两条多肽片段S335-352和S442-458,能与免疫动物血清特异性结合,与使用Bcipep数据库预测B细胞表位的结果相一致。结论鉴定了两个新的SARS病毒S蛋白B细胞表位。
Objective To screen the B cell epitope of SARS virus S protein. Methods BALB / c mice were immunized with SARS-CoV S DNA vaccine to obtain the immune serum of SARS virus S protein. A polypeptide library containing 169 SARS-CoV S proteins with partially overlapping amino acid sequences was synthesized. Peptide fragments were coated on ELISA plates, and immunized mouse sera were used to screen for linear B-cell epitopes of SARS virus S protein by antibody binding assays. The screening results were compared with those predicted using B cell epitope analysis software. Results The two polypeptide fragments of S335-352 and S442-458 of SARS-CoV were screened by overlapping peptide synthesis, which could specifically bind to the serum of immunized animals, which was consistent with the prediction of B cell epitope by Bcipep database. Conclusion Two new SARS virus S protein B cell epitopes were identified.