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Recombinant DNA

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Related Experiment Video

Updated: Jul 5, 2025

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

EPISOMAL VECTORS FOR STABLE PRODUCTION OF RECOMBINANT PROTEINS AND ENGINEERED ANTIBODIES.

Ian Fallahee, Daniel Hawiger

Biorxiv : the Preprint Server for Biology

|January 23, 2024

Summary

A novel plasmid system enables rapid generation of stable Expi293 cell pools for producing recombinant proteins, including bispecific antibodies and antibody-drug conjugates. This versatile method ensures robust protein expression for diverse research and therapeutic applications.

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Last Updated: Jul 5, 2025

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

Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach

Recombinant Protein Expression for Structural Biology in HEK 293F Suspension Cells: A Novel and Accessible Approach

Published on: October 16, 2014

Laboratory Scale Production and Purification of a Therapeutic Antibody

Laboratory Scale Production and Purification of a Therapeutic Antibody

Published on: January 24, 2017

Area of Science:

Biotechnology
Molecular Biology
Protein Engineering

Background:

High demand exists for efficient production of recombinant proteins, especially bispecific antibodies and antibody-drug conjugates for research and therapeutics.
Existing methods for stable cell line generation can be time-consuming and complex.

Approach:

Developed a versatile plasmid system for rapid generation of stable Expi293 cell pools using episomal DNA retention.
Linked protein expression to puromycin resistance via an attenuated internal ribosome entry site for stable cell pool generation.
Utilized split intein-split puromycin selection for co-expression of multiple proteins, enabling bispecific antibodies or modified antibodies.

Key Points:

Achieved stable Expi293 cell pools for consistent production of proteins of interest.
Demonstrated the system's capability to produce bispecific antibody-like molecules with distinct C-terminal modifications.
Generated stable cell pools secreting a sortase A variant (Srt4M) for site-specific conjugation.
Successfully created a site-specific antibody-siRNA conjugate with a defined drug-to-antibody ratio.

Conclusions:

The developed plasmid system offers a simple, robust, and rapid method for stable protein expression.
This technology is broadly applicable for various protein production needs in research and therapeutic development.
Facilitates the creation of complex biologics like antibody-drug conjugates and bispecific antibodies.