Pooled protein immunization for identification of cell surface antigens in Streptococcus sanguinis.
Pooled protein immunization for identification of cell surface antigens in Streptococcus sanguinis.
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DOI：
10.1371/journal.pone.0011666
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发表时间：
2010-07-26
期刊：
影响因子：
3.7
通讯作者：
Xu P
中科院分区：
文献类型：
--
作者：
Ge X;Kitten T;Munro CL;Conrad DH;Xu P
可用的细菌基因组为通过反向疫苗学筛选疫苗提供了机会。需要有效识别表面抗原，以减少该技术的时间和动物成本。我们开发了一种方法，以确定表面抗原的血链球菌，一种常见的感染性心内膜炎致病物种迅速。我们应用生物信息学进行抗原预测，并将抗原合并用于免疫。基于计算机算法和比较基因组分析，选择了47种表面暴露蛋白，包括28种脂蛋白和19种细胞壁锚定蛋白。将这些候选物中的8种蛋白质和2种其它蛋白质合并在一起以免疫兔。抗血清与各蛋白质及S.血细胞亲和层析用于纯化针对9种抗原库组分的抗体。竞争性ELISA和流式细胞仪检测结果表明，这9种蛋白质均能在S.血细胞表面。纯化的抗体具有明显的调理活性。结果表明，免疫与汇集的蛋白质，结合亲和纯化，和全面的免疫学测定可以促进细胞表面抗原鉴定，以打击传染病。
Available bacterial genomes provide opportunities for screening vaccines by reverse vaccinology. Efficient identification of surface antigens is required to reduce time and animal cost in this technology. We developed an approach to identify surface antigens rapidly in Streptococcus sanguinis, a common infective endocarditis causative species. We applied bioinformatics for antigen prediction and pooled antigens for immunization. Forty-seven surface-exposed proteins including 28 lipoproteins and 19 cell wall-anchored proteins were chosen based on computer algorithms and comparative genomic analyses. Eight proteins among these candidates and 2 other proteins were pooled together to immunize rabbits. The antiserum reacted strongly with each protein and with S. sanguinis whole cells. Affinity chromatography was used to purify the antibodies to 9 of the antigen pool components. Competitive ELISA and FACS results indicated that these 9 proteins were exposed on S. sanguinis cell surfaces. The purified antibodies had demonstrable opsonic activity. The results indicate that immunization with pooled proteins, in combination with affinity purification, and comprehensive immunological assays may facilitate cell surface antigen identification to combat infectious diseases.
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影响因子：
3.1
作者：
Brown, JS;Ogunniyi, AD;Paton, JC
通讯作者：
Paton, JC
影响因子：
3.1
作者：
Cole, JN;Ramirez, RD;Walker, MJ
通讯作者：
Walker, MJ
影响因子：
3.1
作者：
Briles, DE;Ades, E;Hollingshead, SK
通讯作者：
Hollingshead, SK
影响因子：
3.1
作者：
Hamilton, Andrea;Robinson, Carl;Harrington, Dean J.
通讯作者：
Harrington, Dean J.
DOI：
10.1073/pnas.092569199
发表时间：
2002-05-14
影响因子：
11.1
作者：
Etz, H;Minh, DB;Meinke, A
通讯作者：
Meinke, A