Micro scale self-interaction chromatography of proteins: A mAb case-study

Area of Science:

• Biopharmaceutical analysis
• Chromatographic techniques
• Protein-protein interactions

Background:

• Self-interaction chromatography (SIC) is a rapid method for determining the second virial coefficient (B22).
• Traditional SIC requires substantial protein quantities (>50 mg), limiting its use in early biotherapeutic screening.
• High protein demand hinders the cost-effective evaluation of novel biotherapeutic candidates.

Purpose of the Study:

• To evaluate a scaled-down self-interaction chromatography (SIC) method.
• To assess the feasibility of using micro-liquid chromatography (LC) columns for SIC.
• To reduce the protein material required for B22 determination in biopharmaceutical development.

Main Methods:

• Evaluation of scaled-down SIC using various micro-LC column dimensions.
• Comparison of results across different column sizes for consistency.
• Quantification of binding (k'), B22, and column volumes.
• Utilized a conventional liquid chromatography system.

Main Results:

• Successful scale-down of SIC was achieved using micro-LC columns.
• Consistent SIC results (k', B22, column volumes) were obtained across different column sizes.
• The scaled-down method requires approximately 10 times less monoclonal antibody (mAb) material compared to conventional SIC.

Conclusions:

• A scaled-down SIC method is effective and reproducible using micro-LC columns.
• This miniaturized approach significantly reduces the protein requirement for B22 determination.
• The method facilitates early-stage screening of biotherapeutic candidates with limited material.