具有oPool+显示的流感血凝素抗体的高通量合成和特异性表征

IF 14.6 1区 医学 Q1 CELL BIOLOGY
Wenhao O. Ouyang, Huibin Lv, Wenkan Liu, Ruipeng Lei, Zongjun Mou, Tossapol Pholcharee, Logan Talmage, Meixuan Tong, Wei Ji, Yiquan Wang, Katrine E. Dailey, Akshita B. Gopal, Danbi Choi, Madison R. Ardagh, Lucia A. Rodriguez, Jenna J. Guthmiller, Xinghong Dai, Nicholas C. Wu
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引用次数: 0

摘要

抗体的发现对于开发治疗方法和疫苗以及了解适应性免疫至关重要。然而,缺乏以高通量方式合成具有确定序列的抗体的方法是抗体发现的主要瓶颈。在这里,我们提出了oPool+显示,这是一种高通量无细胞平台,结合了寡核苷酸池合成和mRNA显示,可以快速构建和表征数百到数千种天然配对抗体。作为概念验证,我们应用oPool+ display通过16个筛选,探测了300多种罕见的流感血凝素特异性抗体对9种血凝素变体的结合特异性。在3至5天的动手时间内进行了5000多次绑定测试,具有进一步的扩展潜力。对两种抗体的后续结构和功能分析显示,人免疫球蛋白基因片段D3-3 (IGHD-3-3)在识别血凝素干细胞方面具有多功能性。总的来说,本研究建立了一个实验平台,不仅可以加速抗体表征,而且可以公正地发现具有相同特异性的抗体之间重复出现的分子特征。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High-throughput synthesis and specificity characterization of natively paired influenza hemagglutinin antibodies with oPool+ display
Antibody discovery is crucial for developing therapeutics and vaccines and for understanding adaptive immunity. However, the lack of approaches to synthesize antibodies with defined sequences in a high-throughput manner represents a major bottleneck in antibody discovery. Here, we present oPool+ display, a high-throughput cell-free platform that combined oligo pool synthesis and mRNA display to rapidly construct and characterize hundreds to thousands of natively paired antibodies in parallel. As a proof of concept, we applied oPool+ display to probe the binding specificity of more than 300 uncommon influenza hemagglutinin-specific antibodies against nine hemagglutinin variants through 16 screens. More than 5000 binding tests were performed in 3 to 5 days of hands-on time with further scaling potential. Follow-up structural and functional analysis of two antibodies revealed the versatility of the human immunoglobulin gene segment D3-3 (IGHD-3-3) in recognizing the hemagglutinin stem. Overall, this study established an experimental platform that not only accelerates antibody characterization but also enables unbiased discovery of recurring molecular signatures among antibodies with the same specificity.
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来源期刊
Science Translational Medicine
Science Translational Medicine CELL BIOLOGY-MEDICINE, RESEARCH & EXPERIMENTAL
CiteScore
26.70
自引率
1.20%
发文量
309
审稿时长
1.7 months
期刊介绍: Science Translational Medicine is an online journal that focuses on publishing research at the intersection of science, engineering, and medicine. The goal of the journal is to promote human health by providing a platform for researchers from various disciplines to communicate their latest advancements in biomedical, translational, and clinical research. The journal aims to address the slow translation of scientific knowledge into effective treatments and health measures. It publishes articles that fill the knowledge gaps between preclinical research and medical applications, with a focus on accelerating the translation of knowledge into new ways of preventing, diagnosing, and treating human diseases. The scope of Science Translational Medicine includes various areas such as cardiovascular disease, immunology/vaccines, metabolism/diabetes/obesity, neuroscience/neurology/psychiatry, cancer, infectious diseases, policy, behavior, bioengineering, chemical genomics/drug discovery, imaging, applied physical sciences, medical nanotechnology, drug delivery, biomarkers, gene therapy/regenerative medicine, toxicology and pharmacokinetics, data mining, cell culture, animal and human studies, medical informatics, and other interdisciplinary approaches to medicine. The target audience of the journal includes researchers and management in academia, government, and the biotechnology and pharmaceutical industries. It is also relevant to physician scientists, regulators, policy makers, investors, business developers, and funding agencies.
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