Pathogenic autoreactive antibodies that may be associated with life-threatening coronavirus disease 2019 (COVID-19) remain to be identified. Here, we show that self-assembled genome-scale libraries of full-length proteins covalently coupled to unique DNA barcodes for analysis by sequencing can be used for the unbiased identification of autoreactive antibodies in plasma samples. By screening 11,076 DNA-barcoded proteins expressed from a sequence-verified human ORFeome library, the method, which we named MIPSA (for Molecular Indexing of Proteins by Self-Assembly), allowed us to detect circulating neutralizing type-I and type-III interferon (IFN) autoantibodies in five plasma samples from 55 patients with life-threatening COVID-19. In addition to identifying neutralizing type-I IFN-α and IFN-ω autoantibodies and other previously known autoreactive antibodies in patient plasma, MIPSA enabled the detection of as yet unidentified neutralizing type-III anti-IFN-λ3 autoantibodies that were not seen in healthy plasma samples or in convalescent plasma from ten non-hospitalized individuals with COVID-19. The low cost and simple workflow of MIPSA will facilitate unbiased high-throughput analyses of protein–antibody, protein–protein and protein–small-molecule interactions.

可能与危及生命的 2019 年冠状病毒病 (COVID-19) 相关的致病性自身反应抗体仍有待鉴定。在这里，我们展示了与独特的 DNA 条形码共价偶联的全长蛋白质的自组装基因组规模文库，用于测序分析，可用于血浆样品中自身反应抗体的公正鉴定。通过筛选从序列验证的人类 ORFeome 文库中表达的 11,076 个 DNA 条形码蛋白质，我们将这种方法命名为 MIPSA（自组装蛋白质分子索引），使我们能够检测循环中和性 I 型和 III 型干扰素(IFN) 自身抗体来自 55 名危及生命的 COVID-19 患者的 5 份血浆样本。除了鉴定患者血浆中的中和性 I 型 IFN-α 和 IFN-ω 自身抗体以及其他先前已知的自身反应性抗体外，MIPSA 还能够检测到尚未鉴定的中和性 III 型抗 IFN-λ3 自身抗体，这些抗体在健康人中未见。来自 10 名未住院的 COVID-19 患者的血浆样本或恢复期血浆。 MIPSA 的低成本和简单的工作流程将有助于对蛋白质-抗体、蛋白质-蛋白质和蛋白质-小分子相互作用进行公正的高通量分析。