Accelerated cell line development using two-color fluorescence activated cell sorting to select highly expressing...

Area of Science:

• Biotechnology
• Biopharmaceutical Development
• Cell Line Engineering

Background:

• Engineered monoclonal antibodies are crucial biopharmaceuticals.
• Developing high-level antibody-expressing mammalian cell lines typically requires 6-12 months.
• Accelerated development strategies are of significant interest.

Purpose of the Study:

• To present a novel method for accelerating the selection of cells expressing recombinant proteins, specifically antibodies.
• To reduce the time and complexity associated with developing stable, high-producing cell lines.

Main Methods:

• Utilized multiparameter fluorescence-activated cell sorting (FACS) combined with dual intracellular autofluorescent reporter proteins.
• Employed a two-color FACS sorting strategy with heavy and light chain-specific fluorescent reporters in Chinese hamster ovary (CHO) cells.
• The method is co-factor-independent and requires minimal sample preparation.

Main Results:

• Achieved cell lines with over a 38-fold increase in antibody production compared to the initial pool after a single FACS round.
• Established high-producing cell lines within 12 weeks of transfection, significantly faster than conventional methods.
• Demonstrated a robust strategy for accelerated clone selection and characterization.

Conclusions:

• The described FACS-based method significantly accelerates the development of antibody-expressing cell lines.
• This approach provides a foundation for predictive modeling of specific productivity using fluorescence data.
• The method offers a robust and efficient strategy for biopharmaceutical cell line development.