Clonal Lineage and Gene Diversity Analysis of Paired Antibody Heavy and Light Chains
Name
nihms-2070482.pdf
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Accepted version
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390.88 KB
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Checksum (MD5)
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Author(s) • • •
Fahad, Ahmed S
Gutiérrez-Gonzalez, Matías F
Madan, Bharat
DeKosky, Brandon J
Date Issued
2025
Journal
Cold Spring Harbor Protocols
Publisher
Cold Spring Harbor Laboratory
Version
Author's final manuscript
Abstract
Antibodies consist of unique variable heavy (VH) and variable light (VL) chains, and both are required to fully characterize an antibody. Methods to detect paired heavy and light chain variable regions (VH:VL) 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 VH:VL 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 VH:VL gene repertoire data to enable efficient antibody discovery.
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Creative Commons Attribution-NonCommercial-ShareAlike
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DOI of Published Version
https://cshprotocols.cshlp.org/content/2025/1/pdb.prot108628.long