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Hybridoma Technology

Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...

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Related Experiment Video

Updated: Jul 17, 2025

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Published on: February 6, 2018

Antibody Batch Cloning.

Rico Ballmann¹, Kai-Thomas Schneider¹, Kristian Daniel Ralph Roth¹

¹Technische Universität Braunschweig, Institute of Biochemistry, Biotechnology and Bioinformatics, Braunschweig, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|September 7, 2023

Summary

This summary is machine-generated.

This study introduces a high-throughput cloning protocol for rapidly screening antibody candidates in bivalent formats. The new method streamlines the production and analysis of bivalent antibody molecules for enhanced characterization.

Keywords:

Batch cloning Parallized cloning Single chain fragment variable (scFv)high-throughput phage display scFv-Fc

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Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Published on: August 22, 2019

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Last Updated: Jul 17, 2025

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

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Published on: February 6, 2018

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

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Published on: June 5, 2015

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Published on: August 22, 2019

Area of Science:

Biotechnology
Immunology
Molecular Biology

Background:

Phage display is crucial for antibody discovery, with initial screening using monovalent single-chain variable fragments (scFv).
Characterizing antibody clones often necessitates bivalent formats like IgG or scFv-Fc fusions for accurate assessment.
Current methods for producing bivalent antibodies involve laborious cloning procedures for V region genes into expression vectors.

Purpose of the Study:

To develop a high-throughput procedure for the rapid screening of antibody candidates in bivalent formats.
To establish a parallelized cloning protocol for selected antibody fragments into mammalian expression vectors.
To enable efficient production and analysis of bivalent antibody molecules in a 96-well plate format.

Main Methods:

A novel high-throughput protocol for parallelized cloning of antibody fragments into a mammalian expression vector.
Utilizing a 96-well plate format for streamlined cloning and subsequent antibody production.
Production and purification of bivalent antibody molecules (e.g., IgG, scFv-Fc) in 96-well plates.

Main Results:

Successful establishment of a high-throughput procedure for antibody screening in bivalent formats.
Demonstrated parallelized cloning of selected antibody fragments into mammalian expression vectors.
Enabled rapid production and purification of bivalent antibody molecules for downstream assays.

Conclusions:

The developed protocol significantly accelerates the screening and characterization of antibody candidates.
This high-throughput method simplifies the production of bivalent antibodies, reducing labor and time.
The 96-well plate format facilitates efficient analysis of antibody clones in bivalent formats for drug discovery and research.