Rapid Buffer and Ligand Screening for Affinity Chromatography by Multiplexed Surface Plasmon Resonance Imaging.

Area of Science:

• Biotechnology
• Biochemistry
• Process Chemistry

Background:

• Affinity chromatography is a key protein purification technique relying on selective ligand binding.
• Optimizing wash and elution conditions is crucial for efficient affinity purification.
• Miniaturization strategies are increasingly used to expedite process development.

Purpose of the Study:

• To evaluate Surface Plasmon Resonance Imaging (SPRi) for screening affinity purification conditions.
• To assess the stability of Protein A ligand under harsh cleaning conditions using SPRi.
• To demonstrate the utility of SPRi in miniaturized process development for recombinant protein purification.

Main Methods:

• Utilized multiplexed Surface Plasmon Resonance Imaging (SPRi) to screen multiple buffer conditions simultaneously.
• Applied SPRi to assess Protein A ligand stability against alkaline conditions.
• Conducted experiments in a miniaturized format, significantly reducing reagent and resin consumption.

Main Results:

• SPRi effectively screened various buffer conditions for wash and elution steps in affinity purification.
• SPRi enabled the assessment of Protein A ligand stability, crucial for resin reuse.
• Miniaturized SPRi experiments reduced resin, protein, and buffer usage by 30-1000 fold compared to conventional methods.
• Development time was decreased from months to days.

Conclusions:

• SPRi is a powerful tool for accelerating the development of affinity purification processes.
• The miniaturized SPRi approach offers substantial savings in resources and time.
• SPRi facilitates early-stage screening for ligand development and recombinant protein production.