High-throughput identification of antibody features for sequence-based epitope prediction

项目总结 由于VDJ重组和体细胞超突变，人类抗体谱高度多样化。VDJ 重组是一种体细胞重组过程，它将抗体的可变区从一个 不同的一组基因片段，称为可变(V)、多样性(D)和连接(J)基因。在…的过程中 作为一种免疫反应，抗体会通过体细胞超突变来增加对其抗原的亲和力。这个 抗体的巨大多样性使人类免疫系统能够通过以下方式保护自己免受各种病原体的侵害 识别广泛的抗原和表位。抗体-抗原的详细分子表征 相互作用对于疫苗和治疗的发展以及对 人类的免疫系统。 抗体的结合特异性和表位由其结构决定，而结构又是 由其氨基酸序列决定。因此，关于结合特异性和表位的信息 抗体是以其氨基酸序列编码的。然而，准确预测抗体的表位 从它们的序列中提取抗体是一项极其困难的任务，因为我们对抗体序列功能的了解 这种关系远远不是全面的。该提案旨在开发一种从图书馆到图书馆的筛选方法，以 以高通量方式描述抗体与抗原的相互作用，重点是甲型流感 血凝素(HA)作为一种概念验证。具体来说，我们将确定HA结合的特异性和 一次实验中数十万种抗体的构象表位。随后，抗体 与HA上不同表位相关的序列特征将被系统地识别。我们进一步 目的利用这些抗体特征从公开提供的抗体库中鉴定HA结合抗体 对数据集进行测序，并预测它们在HA上的表位。虽然这个拟议的项目侧重于流感 哈，我们的方法可以很容易地扩展到任何感兴趣的抗原。这项提议将开辟一种可能性 为基于抗体序列的表位预测提供了新的视角 抗体库。

PROJECT SUMMARY Human antibody repertoire is highly diverse due to VDJ recombination and somatic hypermutation. VDJ recombination is a somatic recombination process that assembles the variable region of an antibody from a diverse set of gene segments, known as variable (V), diversity (D), and joining (J) genes. During the course of an immune response, antibodies will increase affinity to their antigens through somatic hypermutation. The huge diversity of antibodies enables human immune system to confer protection against various pathogens by recognizing a wide range of antigens and epitopes. Detailed molecular characterization of antibody-antigen interaction is crucial to vaccine and therapeutic development, as well as the fundamental understanding of the human immune system. The binding specificity and epitope of an antibody are determined by its structure, which in turn is determined by its amino acid sequence. As a result, information on the binding specificity and epitope of an antibody are encoded in its amino acid sequence. However, accurately predicting the epitopes of antibodies from their sequences is an extremely difficult task because our understanding of antibody sequence-function relationship is far from comprehensive. This proposal aims to develop a library-to-library screening approach to characterize antibody-antigen interaction in a high-throughput manner, with a focus on influenza A hemagglutinin (HA) as a proof-of-concept. Specifically, we will determine the HA-binding specificity and conformational epitope of hundreds of thousands of antibodies in a single experiment. Subsequently, antibody sequence features that are associated with different epitopes on HA will be systematically identified. We further aim to use these antibody features to identify HA-binding antibodies from publicly available antibody repertoire sequencing datasets as well as predict their epitopes on HA. While this proposed project focuses on influenza HA, our approach can be easily extended to any antigen of interest. This proposal will open up the possibility for antibody sequence-based epitope prediction and provides new perspectives to the understanding of human antibody repertoire.