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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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Updated: Jan 11, 2026

Protein Engineering by Yeast Surface Display

Protein Engineering by Yeast Surface Display

Published on: November 29, 2024

Mammalian cell display for antibody engineering.

Mitchell Ho¹, Ira Pastan

¹National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.

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

|March 3, 2009

Summary

This summary is machine-generated.

Mammalian cell display enables antibody engineering for improved affinity. This method successfully isolated high-affinity single-chain variable fragments (scFvs) targeting CD22.

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Last Updated: Jan 11, 2026

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Area of Science:

Biotechnology
Immunology
Molecular Biology

Background:

Traditional antibody engineering relies on microbial display systems like phage, bacteria, and yeast.
Mammalian cell display offers an alternative for engineering antibodies and their fragments.

Purpose of the Study:

To develop and validate a mammalian cell display strategy for antibody affinity maturation.
To isolate high-affinity single-chain variable fragments (scFvs) specific for the CD22 antigen.

Main Methods:

Utilized mammalian cell display, specifically human embryonic kidney 293T (HEK-293T) cells, for transient protein expression.
Employed flow cytometry to enhance screening sensitivity for isolating high-affinity antibodies.
Focused on engineering single-chain variable fragments (scFvs) for increased binding affinity.

Main Results:

Demonstrated the efficacy of mammalian cell display for antibody engineering and affinity maturation.
Successfully isolated high-affinity scFvs targeting the CD22 antigen.
Showcased the potential of this system for engineering other antibody formats like Fabs and whole IgGs.

Conclusions:

Mammalian cell display is a viable and effective platform for antibody engineering, including affinity maturation.
The described strategy using HEK-293T cells and flow cytometry enables the isolation of high-affinity antibodies.
This approach has broad applicability for developing antibodies with enhanced biological functions.