Protein association on multimodal chromatography media

Area of Science:

• Biochemistry and Biophysical Chemistry
• Chromatography and Separation Science
• Protein Science

Background:

• Protein-protein interactions are crucial in biological systems and influence chromatographic behavior.
• Understanding adsorption phenomena on multimodal chromatography media is key for optimizing bioseparations.
• The impact of protein association on adsorption isotherms and elution profiles requires detailed investigation.

Purpose of the Study:

• To investigate protein-protein association on multimodal chromatography resins.
• To elucidate the adsorption patterns of alpha-lactalbumin (aLaCa, aLa) and bovine serum albumin (BSA) mixtures.
• To examine the adsorption of monoclonal antibody (mAb) and lysozyme (LYZ) mixtures on multimodal media.

Main Methods:

• Multimodal anion-exchange and cation-exchange chromatography were employed.
• Single-component and binary mixtures of proteins were analyzed under varying pH and buffer conditions.
• A mechanistic model incorporating protein association and exclusion was utilized for quantification.

Main Results:

• Alpha-lactalbumin showed concentration-dependent adsorption changes, with higher adsorption at high loadings.
• Bovine serum albumin adsorption was enhanced in binary mixtures with alpha-lactalbumin.
• Monoclonal antibody adsorption was enhanced in the presence of lysozyme, indicating protein association effects.

Conclusions:

• Protein-protein association in the adsorbed phase significantly affects adsorption behavior on multimodal chromatography resins.
• The findings provide insights into optimizing chromatographic separations by considering protein interactions.
• The mechanistic model effectively quantifies the influence of protein association and exclusion on adsorption.