Monitoring an antibody affinity chromatography with a label-free optical biosensor technique

Area of Science:

• Biotechnology
• Analytical Chemistry
• Immunology

Background:

• Polyclonal antibody production requires purification, typically via affinity chromatography.
• Monitoring antibody quantity and quality during purification is crucial for downstream applications.
• Existing methods like ELISA are time-consuming and lack real-time process insights.

Purpose of the Study:

• To develop and validate a biosensor system for real-time monitoring of polyclonal antibody purification.
• To assess antibody quantity, quality (affinity, kinetics), and active antibody to protein concentration ratio.
• To enable rapid in-process monitoring of affinity chromatography for optimal fraction selection.

Main Methods:

• Utilized a biosensor system based on reflectometric interference spectroscopy (RIfS).
• Applied the biosensor to monitor antibody purification during affinity chromatography.
• Developed methods for accurate determination of kinetic and affinity constants.

Main Results:

• The RIfS biosensor system provided rapid and automated monitoring of antibody purification.
• Accurate quantification of antibody affinity and kinetic constants was achieved.
• The system enabled determination of the active antibody to protein concentration ratio.
• A new rapid method for in-process monitoring of affinity chromatography was demonstrated.

Conclusions:

• Reflectometric interference spectroscopy (RIfS) biosensors offer a significant advancement for monitoring antibody purification.
• Real-time data on antibody quantity and quality allows for informed selection of antibody fractions.
• This automated approach enhances efficiency and quality control in polyclonal antibody production.