Immunology Division, Department of Pathology, Institute for Cell Engineering, Johns Hopkins University, Baltimore, MD, USA.
Johns Hopkins University Applied Physics Laboratory, Laurel, MD, USA.
EBioMedicine. 2021 Sep;71:103506. doi: 10.1016/j.ebiom.2021.103506. Epub 2021 Sep 1.
Post-translational modifications (PTMs) on proteins can be targeted by antibodies associated with autoimmunity. Despite a growing appreciation for their intrinsic role in disease, there is a lack of highly multiplexed serological assays to characterize the fine specificities of PTM-directed autoantibodies.
In this study, we used the programmable phage display technology, Phage ImmunoPrecipitation Sequencing (PhIP-Seq), to profile rheumatoid arthritis (RA) associated anti-citrullinated protein antibody (ACPA) reactivities.
Using both unmodified and peptidylarginine deiminase (PAD)-modified phage display libraries consisting of ~250,000 overlapping 90 amino acid peptide tiles spanning the human proteome, PTM PhIP-Seq robustly identified antibodies to citrulline-dependent epitopes.
PTM PhIP-Seq was used to quantify key differences among RA patients, including PAD isoform specific ACPA profiles, and thus represents a powerful tool for proteome-scale antibody-binding analyses.
This research is based upon work supported in part by the Office of the Director of National Intelligence (ODNI), Intelligence Advanced Research Projects Activity (IARPA). The views and conclusions contained herein are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of ODNI, IARPA, or the US Government. The US Government is authorized to reproduce and distribute reprints for governmental purposes notwithstanding any copyright annotation therein. This study was made possible by a National Institute of General Medical Sciences (NIGMS) grant R01 GM136724 (HBL). MFK was supported by the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) grant T32AR048522. ED was supported by the Rheumatology Research Foundation.
蛋白质的翻译后修饰（PTMs）可以被与自身免疫相关的抗体靶向。尽管人们越来越认识到它们在疾病中的固有作用，但缺乏高度多重化的血清学检测方法来描述 PTM 定向自身抗体的精细特异性。
在这项研究中，我们使用可编程噬菌体展示技术，噬菌体免疫沉淀测序（PhIP-Seq），来描绘类风湿关节炎（RA）相关的抗瓜氨酸化蛋白抗体（ACPA）反应性。
使用未修饰和肽基精氨酸脱亚氨酶（PAD）修饰的噬菌体展示文库，文库由约 250000 个重叠的 90 个氨基酸肽组成，覆盖人类蛋白质组，PTM PhIP-Seq 可有效地鉴定依赖瓜氨酸的表位的抗体。
PTM PhIP-Seq 用于量化 RA 患者之间的关键差异，包括 PAD 同工型特异性 ACPA 谱，因此代表了一种用于蛋白质组范围抗体结合分析的强大工具。
这项研究的部分工作得到了美国国家情报总监办公室（ODNI）、情报高级研究项目活动（IARPA）的支持。本文中的观点和结论仅代表作者的观点，不应被解释为代表 ODNI、IARPA 或美国政府的官方政策，无论是明确表达的还是暗示的。美国政府有权复制和分发本研究报告的副本，用于政府目的，尽管其中有版权注释。这项研究得到了国立普通医学科学研究所（NIGMS）R01 GM136724 （HBL）资助的支持。MFK 得到了国立关节炎和肌肉骨骼及皮肤病研究所（NIAMS）T32AR048522 资助的支持。ED 得到了风湿病研究基金会的支持。
