成对抗体重链和轻链的克隆谱系和基因多样性分析

Cold Spring Harbor protocols Pub Date : 2024-11-25 DOI:10.1101/pdb.prot108628

Ahmed S Fahad, Matías F Gutiérrez-Gonzalez, Bharat Madan, Brandon J DeKosky

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引用次数: 0

摘要

抗体由独特的可变重链（VH）和可变轻链（VL）组成，要全面描述抗体的特性，这两条链都必不可少。最近，利用高通量测序（HTS）检测成对的重链和轻链可变区（VH:VL）的方法实现了对完整的功能性抗体反应的大规模分析。在此，我们介绍一种 HTS 计算管道，用于分析成对的 VH:VL 抗体序列，并获得免疫多样性景观的综合概况，包括基因使用、抗体异型和克隆谱系分析。该方案使用 Illumina MiSeq 2 × 300-bp 测序数据，并集成了多种不同的计算工具，可灵活分析成对的 VH:VL 基因库数据，从而实现高效的抗体发现。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Clonal Lineage and Gene Diversity Analysis of Paired Antibody Heavy and Light Chains.

Antibodies consist of unique variable heavy (V_H) and variable light (V_L) chains, and both are required to fully characterize an antibody. Methods to detect paired heavy and light chain variable regions (V_H:V_L) using high-throughput sequencing (HTS) have recently enabled large-scale analysis of complete functional antibody responses. Here, we describe an HTS computational pipeline to analyze paired V_H:V_L antibody sequences and obtain a comprehensive profile of immune diversity landscapes, including gene usage, antibody isotypes, and clonal lineage analysis. This protocol uses Illumina MiSeq 2 × 300-bp sequencing data and integrates with several different computational tools for flexible analyses of paired V_H:V_L gene repertoire data to enable efficient antibody discovery.

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来源期刊

Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)

CiteScore

3.00

自引率

0.00%

发文量

163

期刊介绍： Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.