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Updated: Mar 16, 2026

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

Published on: April 7, 2023

High-quality full-length immunoglobulin profiling with unique molecular barcoding.

M A Turchaninova^1,2,3, A Davydov², O V Britanova^1,3

¹Shemiakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Science, Moscow, Russia.

Nature Protocols

|August 5, 2016

Summary

This summary is machine-generated.

This study introduces a new protocol for high-throughput sequencing of immunoglobulin (IG) repertoires. It uses unique molecular identifiers (UMIs) for accurate, full-length profiling of human and mouse antibody genes.

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Published on: December 28, 2015

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Last Updated: Mar 16, 2026

Multiplexed Barcoding Image Analysis for Immunoprofiling and Spatial Mapping Characterization in the Single-Cell Analysis of Paraffin Tissue Samples

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VDJ-Seq: Deep Sequencing Analysis of Rearranged Immunoglobulin Heavy Chain Gene to Reveal Clonal Evolution Patterns of B Cell Lymphoma

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Published on: December 28, 2015

Area of Science:

Immunology
Genomics
Bioinformatics

Background:

High-throughput sequencing of immunoglobulin (IG) repertoires is crucial for understanding immune responses but faces challenges with accuracy and length.
Existing methods struggle with errors introduced during PCR amplification and sequencing, limiting comprehensive repertoire analysis.

Purpose of the Study:

To develop a robust protocol for full-length profiling of human and mouse IG repertoires.
To enhance accuracy and sequence length in IG repertoire sequencing through error correction methods.

Main Methods:

Implementation of unique molecular identifiers (UMIs) during cDNA synthesis to correct for PCR and sequencing errors.
Utilizing asymmetric 400+100-nt paired-end Illumina sequencing.
Employing the MIGEC software for UMI-based assembly of sequences.

Main Results:

Achieved near error-free sequencing of up to 750-nt lengths, covering variable and partial constant regions of IG chains.
The protocol enables accurate full-length profiling of both human and mouse IG repertoires.
Demonstrated applicability of the sequencing approach beyond immune repertoire studies.

Conclusions:

The developed protocol offers a robust and accurate method for high-throughput IG repertoire profiling.
This approach significantly improves the quality and length of sequence data, providing deeper insights into antibody diversity.
The methodology is adaptable for various sequencing applications requiring high accuracy and long reads.