Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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, polyethylene glycol...

Immunoprecipitation

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

A hexamerization-enhanced, Fc-silenced agonistic CD27 antibody amplifies T-cell effector functions as single agent and in combination with PD-1 blockade.

Scientific reports·2025

Same author

Logic-gated antibody pairs that selectively act on cells co-expressing two antigens.

Nature biotechnology·2022

Same author

Biophysical Characterization and Stability of Modified IgG1 Antibodies with Different Hexamerization Propensities.

Journal of pharmaceutical sciences·2022

Same author

Staphylococcal protein A inhibits complement activation by interfering with IgG hexamer formation.

Proceedings of the National Academy of Sciences of the United States of America·2021

Same author

Releasing Nonperipheral Subunits from Protein Complexes in the Gas Phase.

Analytical chemistry·2020

Same author

Dual Epitope Targeting and Enhanced Hexamerization by DR5 Antibodies as a Novel Approach to Induce Potent Antitumor Activity Through DR5 Agonism.

Molecular cancer therapeutics·2020

Search research articles

Related Experiment Video

Updated: May 9, 2026

Laboratory Scale Production and Purification of a Therapeutic Antibody

Laboratory Scale Production and Purification of a Therapeutic Antibody

Published on: January 24, 2017

A simple, robust and highly efficient transient expression system for producing antibodies.

Tom Vink¹, Maroeska Oudshoorn-Dickmann¹, Marcel Roza¹

¹Genmab, Yalelaan 60, 3584 CM, Utrecht, The Netherlands.

Methods (San Diego, Calif.)

|July 23, 2013

Summary

This summary is machine-generated.

This study presents an optimized mammalian transient expression system for rapid, high-yield antibody production. The system achieves up to 400mg/L of native antibodies in under a week using HEK-293F cells.

Keywords:

293Fectin 293fectin™Antibody CMV Cell cycle DoE Experimental design FreeStyle™ 293 expression medium FreeStyle™ 293-F HC HEK-293 HEK-293F HEK293 LC Medium Optimem SV40 large T antigen SVLT Transient expression antibody heavy chain antibody light chain cyclin-dependent kinase inhibitor 1 cyclin-dependent kinase inhibitor 1B cytomegalo virus design of experiments human embryonic kidney 293 cell line opti-MEM® I reduced-serum medium p21 p27 the large T antigen of the SV40 virus

More Related Videos

A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

Published on: July 31, 2012

Controllable Ion Channel Expression through Inducible Transient Transfection

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

Related Experiment Videos

Last Updated: May 9, 2026

Laboratory Scale Production and Purification of a Therapeutic Antibody

Laboratory Scale Production and Purification of a Therapeutic Antibody

Published on: January 24, 2017

A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells

Published on: July 31, 2012

Controllable Ion Channel Expression through Inducible Transient Transfection

Controllable Ion Channel Expression through Inducible Transient Transfection

Published on: February 17, 2017

Area of Science:

Biotechnology
Molecular Biology
Protein Expression

Background:

Mammalian transient expression systems are preferred for antibody production due to speed, yield, and posttranslational modifications.
Existing systems have limitations in terms of maximal achievable yield and production time.

Purpose of the Study:

To develop and optimize a mammalian transient expression system for high-yield, rapid production of native secreted antibodies.
To identify key components and conditions for maximizing antibody expression levels.

Main Methods:

Utilized the FreeStyle™ 293-F (HEK-293F) suspension cell line for expression.
Optimized gene transfer, expression vector, and co-transfection with SV40 large T antigen and p21/p27 plasmids.
Employed experimental design techniques for component optimization.
Scaled expression from 0.1 mL to 1.2 L in shaker flasks using animal component-free medium.

Main Results:

Achieved antibody yields up to 400 mg/L in less than a week.
Successfully produced various antibody isotypes (IgG1, 2, 3, 4) and Fab fragments from different species.
Demonstrated scalability and consistency in production volumes.

Conclusions:

The optimized system provides a robust and efficient method for producing research quantities of native antibodies.
This system offers a significant improvement over existing transient expression methods for antibody manufacturing.
The scalable, rapid, and high-yield nature makes it suitable for diverse antibody production needs.