Integrated system for temperature-controlled fast protein liquid chromatography. II. Optimized adsorbents and 'single...

Area of Science:

• Biochemistry
• Chromatography
• Materials Science

Background:

• Traditional chromatography methods face limitations in efficiency and selectivity for complex biopurification.
• Thermoresponsive polymers offer tunable properties for dynamic control in separation processes.
• Developing advanced chromatography systems is crucial for isolating valuable biomolecules.

Purpose of the Study:

• To develop and evaluate a novel temperature-controlled chromatography system (TCZR).
• To synthesize and characterize thermoresponsive cation exchangers (thermoCEX) with tunable properties.
• To assess the performance of the TCZR system for lactoferrin binding, concentration, and separation from bovine serum albumin.

Main Methods:

• Synthesis of nine copolymer-grafted thermoresponsive cation exchangers (thermoCEX) using different agarose supports.
• Batch binding tests to determine maximum binding capacity (qmax) and thermoresponsiveness.
• Chromatographic experiments using the travelling cooling zone reactor (TCZR) with lactoferrin and lactoferrin/bovine serum albumin mixtures.

Main Results:

• ThermoCEX supports with optimal thermoresponsiveness and binding capacity were identified (A2, B2, C1).
• The TCZR system demonstrated intermittent concentration of lactoferrin into sharp, uniform peaks.
• Successful separation of lactoferrin from bovine serum albumin was achieved, with minor impacts on separation efficiency.

Conclusions:

• The developed thermoresponsive cation exchangers (thermoCEX) and TCZR system show significant promise for efficient protein purification.
• The system enables dynamic control over binding and elution through temperature modulation, enhancing concentration factors.
• Further optimization is needed to mitigate potential matrix fouling during complex mixture separations.