Development of versatile affinity-based system for one step purification process: Case of Group A Streptococcus...

Area of Science:

• Biotechnology
• Protein Engineering
• Vaccine Development

Background:

• Affinity capture is crucial for antibody purification but limited for other biologics like vaccines due to a lack of suitable materials.
• Current purification methods for biologics often involve multiple steps, increasing complexity and reducing efficiency.

Purpose of the Study:

• To introduce a novel custom affinity system for broad application in biologics purification beyond antibodies.
• To develop a streamlined, one-step purification process for a streptolysin O (SLO) vaccine candidate.

Main Methods:

• Developed custom ligands based on the Nanofitin protein scaffold, selected for target specificity.
• Integrated Nanofitins with Eshmuno® chromatography medium to create a novel affinity medium.
• Validated the system's scalability across various column sizes, from process development to pilot runs.

Main Results:

• Achieved a high purity product (above 90%) in a single purification step.
• Demonstrated significant improvements over a three-step process: reduced time (3 to 1 step), smaller footprint, and increased product yield (0.31 g vs. 0.04 g SLO per kg harvest broth).
• Confirmed scalability and robustness of the affinity medium.

Conclusions:

• The custom Nanofitin-Eshmuno® affinity system offers an efficient platform for biologics purification, including vaccines.
• This approach simplifies downstream processing, reduces manufacturing costs, and has the potential for wide application in biologic manufacturing.
• Enables efficient production of vaccine candidates like SLO.