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Updated: Jun 6, 2025

VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma
Published on: December 28, 2015
Clonal Lineage and Gene Diversity Analysis of Paired Antibody Heavy and Light Chains
Ahmed S Fahad1,2, Matías F Gutiérrez-Gonzalez1,2, Bharat Madan1
1The Ragon Institute of Massachusetts General Hospital, Massachusetts Institute of Technology, and Harvard University, Cambridge, Massachusetts 02139, USA.
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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