Intracellular Antibody Levels Predict Secretion Efficiency in HEK293T Cells Expressing a Chimeric Anti-CD99 Antibody...

Area of Science:

• Biotechnology
• Molecular Biology
• Immunology

Background:

• Recombinant antibody production is crucial for various applications.
• Single-expression plasmids offer consistent co-expression of antibody chains.
• Predicting antibody secretion efficiency from intracellular levels is challenging.

Purpose of the Study:

• To investigate the correlation between intracellular antibody expression and secretion efficiency.
• To assess the utility of intracellular expression levels for selecting high-producing cell lines.
• To optimize recombinant antibody production using single-plasmid systems.

Main Methods:

• Constructed a single-expression plasmid for a chimeric anti-CD99 antibody (ChAbMT99/1).
• Established stably expressing HEK293T mini-pools via zeocin selection.
• Quantified intracellular antibody levels using flow cytometry and extracellular secretion using ELISA and flow cytometry.

Main Results:

• Demonstrated a strong positive correlation between intracellular antibody expression and secretion levels.
• Higher intracellular fluorescence intensity predicted greater antibody yields in mini-pools.
• Intracellular expression levels reliably predicted secretion efficiency in single-plasmid systems.

Conclusions:

• Intracellular antibody expression is a reliable indicator of secretion efficiency in single-plasmid systems.
• This correlation facilitates efficient selection of high-producing cell populations.
• Optimized selection strategies can enhance large-scale recombinant antibody production.