Robust Diagnostics, LLC, Chandler, Arizona 85226, United States of America.
Anal Chem. 2024 Sep 10;96(36):14524-14530. doi: 10.1021/acs.analchem.4c02806. Epub 2024 Aug 29.
Immunodominant epitope discovery platforms play an important role in identifying novel biomarkers for effective immunotherapies and diagnostics. Methods to analyze the B-cell repertoire have been improved both experimentally and computationally. We developed an enhanced peptide microarray platform to discover and subsequently screen immunodominant epitopes. We utilized SARS-Cov-2 IgG positive and negative samples as a proof-of-concept to demonstrate the power of these improved peptide microarrays. The method identified significantly discriminant epitopes that classify positive and negative samples with good performance both as single peptides and in combination. We provide the assay conditions and parameters that justify the use of peptide microarrays in the selection of high-affinity epitopes, and we directly compare peptide performance against proteins. The results suggest that this platform can be used to confidently identify immunodominant antiviral epitopes while also serving as a useful tool for high-volume screening.
免疫优势表位发现平台在鉴定有效的免疫疗法和诊断学的新型生物标志物方面发挥着重要作用。分析 B 细胞库的方法在实验和计算上都得到了改进。我们开发了一种增强型肽微阵列平台,用于发现和随后筛选免疫优势表位。我们利用 SARS-CoV-2 IgG 阳性和阴性样本作为概念验证,展示了这些改进型肽微阵列的强大功能。该方法鉴定了具有显著区分能力的表位,这些表位可以作为单个肽和组合使用,很好地区分阳性和阴性样本。我们提供了检测条件和参数,证明了在选择高亲和力表位时使用肽微阵列的合理性,并且我们直接比较了肽与蛋白质的性能。结果表明,该平台可用于自信地鉴定免疫优势抗病毒表位,同时也可作为高通量筛选的有用工具。