Purification and characterization of rabbit polyclonal immunoglobulin G using DEAE ion-exchange chromatography and dynamic light scattering

Weak anion-exchange chromatography (AEC) provides a mild alternative to ammonium sulfate (AS) precipitation for antibody purification, but its mechanistic behavior in complex serum systems remains unclear. This study evaluates pH-dependent DEAE flow-through purification of rabbit anti-HBcAg IgG and compares it with AS precipitation. At pH 8.0, DEAE chromatography achieved 94% recovery and 83% purity, consistent with reduced IgG–resin interaction and selective retention of anionic serum proteins. Conductivity profiling showed a 69% reduction in ionic strength relative to AS-treated samples, correlating with improved structural homogeneity. Dynamic light scattering revealed a monomodal ∼10 nm hydrodynamic diameter for DEAE-purified IgG, whereas AS precipitation generated multimodal distributions indicative of aggregation. These results clarify how pH, ionic strength, and protein–protein interactions govern IgG behavior during weak AEC. The findings position DEAE chromatography as a gentle, aggregation-minimizing, and scalable method for producing high-quality polyclonal antibodies for diagnostic applications.

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