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Related Experiment Video

Updated: Jan 23, 2026

Generation of Monoclonal Antibodies Against Natural Products

Generation of Monoclonal Antibodies Against Natural Products

Published on: April 6, 2019

A simplified workflow for monoclonal antibody sequencing.

Lena Meyer¹, Tomás López², Rafaela Espinosa²

¹Department of Biomolecular Engineering, University of California Santa Cruz, Santa Cruz, California, United States of America.

|June 25, 2019

Summary

This summary is machine-generated.

This study introduces a streamlined RT-PCR method for sequencing antibody variable regions, simplifying monoclonal antibody characterization. The approach avoids degenerate primers, enabling accurate sequencing for recombinant antibody production.

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Last Updated: Jan 23, 2026

Generation of Monoclonal Antibodies Against Natural Products

Generation of Monoclonal Antibodies Against Natural Products

Published on: April 6, 2019

Generation of Murine Monoclonal Antibodies by Hybridoma Technology

Generation of Murine Monoclonal Antibodies by Hybridoma Technology

Published on: January 2, 2017

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens

Passive Administration of Monoclonal Antibodies Against H. capsulatum and Others Fungal Pathogens

Published on: February 14, 2011

Area of Science:

Immunology
Molecular Biology
Biotechnology

Background:

Antibody variable region sequencing is crucial for antibody engineering.
Conventional methods for monoclonal antibody cDNA amplification are complex and require degenerate primers due to sequence diversity.

Purpose of the Study:

To develop a simplified, user-friendly, and cost-effective workflow for amplifying and sequencing IgG antibody variable regions.
To enable accurate amino acid sequence determination for recombinant antibody production.

Main Methods:

A specialized reverse transcription PCR (RT-PCR) workflow using a template-switch oligonucleotide.
Three separate reactions for kappa, lambda, and heavy chain transcripts.
Sanger sequencing of amplified antibody variable regions.

Main Results:

Successfully sequenced variable regions of five mouse monoclonal IgG antibodies.
Enabled design of chimeric mouse/human antibody expression plasmids for recombinant production.
Confirmed accuracy through high-affinity antigen binding of recombinant antibodies and applicability to human antibodies.

Conclusions:

The developed method is highly accurate, user-friendly, and cost-effective for monoclonal antibody sequencing.
This workflow facilitates efficient recombinant antibody production and characterization.
The method is applicable to both mouse and human IgG antibodies.