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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: Jun 28, 2025

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells

Rapid Antibody Glycoengineering in Chinese Hamster Ovary Cells

Published on: June 2, 2022

A systemic approach to identifying sequence frameworks that decrease mAb production in a transient Chinese hamster

Alana C Szkodny¹, Kelvin H Lee¹

¹Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.

Biotechnology Progress

|April 12, 2024

Summary

Antibody sequence directly impacts protein expression levels. This study identified specific mutations decreasing antibody expression in CHO cells, aiding in selecting better drug candidates early.

Keywords:

CHO cells developability difficult‐to‐express monoclonal antibodies transient expression

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A High-throughput Automated Platform for the Development of Manufacturing Cell Lines for Protein Therapeutics

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A High-throughput Automated Platform for the Development of Manufacturing Cell Lines for Protein Therapeutics

A High-throughput Automated Platform for the Development of Manufacturing Cell Lines for Protein Therapeutics

Published on: September 22, 2011

Area of Science:

Biotechnology
Protein Engineering
Biopharmaceutical Manufacturing

Background:

Monoclonal antibodies (mAbs) are engineered for clinical efficacy, but their "developability" (e.g., expression levels) is often overlooked before candidate selection.
Poor expression, linked to sequence variations, necessitates costly manufacturing process improvements.
Difficult-to-express (DTE) mAbs exhibit post-translational bottlenecks in folding, assembly, and secretion, but findings are hard to generalize.

Purpose of the Study:

To systematically investigate the impact of sequence variation on mAb expression at scale under industrially relevant conditions.
To identify specific sequence determinants affecting mAb expression in Chinese hamster ovary (CHO) cells.
To develop a workflow for improving mAb candidate selection and reducing process development timelines.

Main Methods:

Engineered a library of monoclonal antibody (mAb) variants with sequence modifications.
Evaluated transient expression titers of these variants in Chinese hamster ovary (CHO) cells.
Analyzed the relationship between specific mutations, residue properties (e.g., hydrophobicity), and expression levels.

Main Results:

Identified 91 mutations that significantly decreased transient expression of an IgG1κ antibody.
Found that mutations at inaccessible residues, particularly those reducing hydrophobicity, are detrimental to high antibody expression.
Demonstrated a strong correlation between primary amino acid sequence and antibody expression titers, with variations causing up to tenfold differences.

Conclusions:

Sequence variations significantly influence mAb expression, presenting a critical developability characteristic.
Understanding sequence-expression relationships allows for early identification of problematic mAbs.
This systematic approach enhances antibody candidate selection and streamlines bioprocess development.