Lorenz Peter, Kreutzer Michael, Zerweck Johannes, Schutkowski Mike, Thiesen Hans-Jürgen
Institute of Immunology, University of Rostock, Schillingallee 70, D-18055 Rostock, Germany.
Methods Mol Biol. 2009;524:247-58. doi: 10.1007/978-1-59745-450-6_18.
High density peptide microarray technologies can be applied in experimental medicine in general and in clinical immunology in particular. Laboratory diagnostics of autoimmune diseases strongly rely on screening human sera for antibodies against known autoantigens. These assays are still difficult to standardize and quantify. Typically, the results are presented as antibody titers within an assay system. Most assays use recombinant or purified autoantigens that are difficult to obtain and require great efforts of quality control. Here we describe a method to obtain patterns of epitope signatures with peptide microarrays from patients suffering from autoimmune diseases in comparison with healthy controls. One of the final aims will be to define subsets of peptides indicative for marker autoantibodies of autoimmune diseases. Finally, informative epitopes can be used for immunopurifying epitope-specific autoantibodies. Eventually, these antibodies can be further characterized on peptide microarrays displaying mutated epitopes obtained by scanning mutagenesis. Any disease or physiological status that affect humoral immune responses such as autoantibodies in oncology or responses to infections or vaccinations can be monitored.
高密度肽微阵列技术一般可应用于实验医学，尤其适用于临床免疫学。自身免疫性疾病的实验室诊断严重依赖于筛查人血清中针对已知自身抗原的抗体。这些检测方法仍难以标准化和定量。通常，结果以检测系统内的抗体滴度表示。大多数检测使用难以获得的重组或纯化自身抗原，并且需要大量的质量控制工作。在此，我们描述了一种方法，用于从自身免疫性疾病患者与健康对照者中获得肽微阵列的表位特征模式。最终目标之一是定义指示自身免疫性疾病标记自身抗体的肽亚组。最后，信息丰富的表位可用于免疫纯化表位特异性自身抗体。最终，这些抗体可在展示通过扫描诱变获得的突变表位的肽微阵列上进一步表征。任何影响体液免疫反应的疾病或生理状态，如肿瘤学中的自身抗体或对感染或疫苗接种的反应，都可以进行监测。
