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Updated: Jun 20, 2026
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Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor
Published on: February 1, 2015
A GFP-based method facilitates clonal selection of transfected CHO cells
Denise Freimark1, Valèrie Jèrôme, Ruth Freitag
1Chair for Process Biotechnology University of Bayreuth, Germany.
Biotechnology Journal
|September 4, 2009
Summary
This study introduces a novel method for selecting high-producing cell clones using enhanced green fluorescent protein (EGFP) co-expression. This technique simplifies recombinant protein production by correlating EGFP levels with therapeutic protein expression in CHO cells.
Area of Science:
- Biotechnology
- Molecular Biology
- Cell Biology
Background:
- Developing cell lines for recombinant protein production requires identifying high-expressing clones.
- Current methods for clonal selection can be complex and require specialized equipment.
Purpose of the Study:
- To present a new method for clonal selection based on enhanced green fluorescent protein (EGFP) co-expression.
- To provide an alternative to cell sorting methods for identifying high-producer clones.
Main Methods:
- Co-expression of EGFP and the protein of interest using an internal ribosome entry site (IRES) construct.
- Independent translation of both proteins from a single mRNA transcript.
- Selection of clones in standard 96-well cell culture plates based on EGFP fluorescence.
Main Results:
- EGFP expression levels directly correlate with the expression levels of the therapeutic protein.
- Demonstrated robustness and performance of the method using recombinant growth factors in CHO cells.
- Successful clonal selection using standard laboratory equipment.
Conclusions:
- The EGFP co-expression method offers a simplified and accessible approach for identifying high-producer clones.
- This technique is a viable alternative to cell sorting for recombinant protein production.
- Facilitates efficient development of cell lines for biopharmaceutical manufacturing.
