Two-Step Purification of Low-Expressing Recombinant Exoprotein A Using Mixed-Mode Chromatography

This case study demonstrates an efficient purification workflow for a low-expressing recombinant protein. Multiple chromatography resins were screened for the capture step of this recombinant protein, with goals to minimize fermentation dilution and increase purity and recovery to >75%. Optimal wash and elution conditions were determined using a design of experiments (DOE) approach. An elution buffer containing NaCl and MgCl₂ was found to purify the target molecule with high recovery and purity.

Overview

• Importance of screening multiple resins
• DOE for optimizing purification and elution conditions
• Evaluation of modifiers for enhancing purity and recovery