寨卡病毒特异性诊断表位的发现
Zika Virus Specific Diagnostic Epitope Discovery.
作者信息
Sabalza Maite, Barber Cheryl A, Abrams William R, Montagna Richard, Malamud Daniel
机构信息
Department of Basic Sciences and Craniofacial Development, New York University College of Dentistry;
Department of Basic Sciences and Craniofacial Development, New York University College of Dentistry.
出版信息
J Vis Exp. 2017 Dec 12(130):56784. doi: 10.3791/56784.
High-density peptide microarrays allow screening of more than six thousand peptides on a single standard microscopy slide. This method can be applied for drug discovery, therapeutic target identification, and developing of diagnostics. Here, we present a protocol to discover specific Zika virus (ZIKV) diagnostic peptides using a high-density peptide microarray. A human serum sample validated for ZIKV infection was incubated with a high-density peptide microarray containing the entire ZIKV protein translated into 3,423 unique 15 linear amino acid (aa) residues with a 14-aa residue overlap printed in duplicate. Staining with different secondary antibodies within the same array, we detected peptides that bind to Immunoglobulin M (IgM) and Immunoglobulin G (IgG) antibodies present in serum. These peptides were selected for further validation experiments. In this protocol, we describe the strategy followed to design, process, and analyze a high-density peptide microarray.
高密度肽微阵列能够在一张标准显微镜载玻片上筛选六千多种肽。该方法可应用于药物发现、治疗靶点鉴定以及诊断方法开发。在此，我们展示了一种使用高密度肽微阵列发现特定寨卡病毒（ZIKV）诊断肽的方案。将一份经寨卡病毒感染验证的人血清样本与一个高密度肽微阵列孵育，该微阵列包含整个寨卡病毒蛋白，其被翻译成3423个独特的15个线性氨基酸（aa）残基，且有14个氨基酸残基重叠，以一式两份打印。在同一阵列中用不同的二抗进行染色，我们检测到了与血清中存在的免疫球蛋白M（IgM）和免疫球蛋白G（IgG）抗体结合的肽。这些肽被选用于进一步的验证实验。在本方案中，我们描述了设计、处理和分析高密度肽微阵列所遵循的策略。
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