FIG 4.

Purification via size exclusion chromatography (SEC) impacts the neutralization potency of D5_AR and reveals possible aggregation. (A and B) UV traces from SEC, performed after affinity chromatography, reveal possible protein aggregates (circled in red) for both D5 and D5_AR IgG. (A) The potential aggregate peak for D5_AR accounts for 11% of the total protein, while the major IgG fraction accounts for 83%. (B) Similarly, the aggregate fraction for D5 accounts for 8.2% of the total protein compared to the 90% of the major D5 IgG fraction. (C) After SEC purification, D5_AR IgG preparations demonstrated enhanced neutralization activity compared to that of a prep that was only purified via affinity chromatography. Each data point represents the mean percent infection with standard error of the mean (n = 3). (D) SEC-purified D5_AR IgG sample preps stored in 10% glycerol-1× PBS solution at −20°C retained similar neutralization activity after being thawed. Each data point represents the mean percent infection with standard error of the mean (n = 3).