Anion-exchange chromatography is a useful and effective tool for adenoviral vectors purification. However, due to the different functional groups and matrices, both binding capacity and resolution of most AEC resins are different. In this study, four different AEC resins are compared by the binding capacity, resolution and recovery. Using Fractogel TMAE as an adsorbent to purify adenoviral vectors has obvious advantages over the other resins, namely (1) dynamic binding capacity is higher than other resins; (2) unprecedented sharpness (1,570,000 ± 250,000) and symmetry of adenoviral vectors peak (1.67 ± 0.06); (3) higher resolution with other contaminants (2.16 ± 0.08); (4) no enzymatic treatment; (5) the recovery can reach 75%; (6) the purity is higher and the total virion to infectious particle ratios can reach 18.9. In the present work, we confirmed the possibility of purifying pharmaceutical-grade adenoviral vectors by AEC.Graphical abstract
Evaluated four anion-exchange chromatography with different functional groups and matrix from column performances (column efficiency, peak asymmetry and resolution), dynamic binding capacity, recovery and purity.