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. 2021 Jul 30;13(8):1503. doi: 10.3390/v13081503

Figure 4.

Figure 4

Capacity and optimal elution from 5 mL Heparin HiTrap columns. HepAD38 cells were cultivated in a HYPERflask and different volumes of HBV-rich supernatant were loaded onto a single 5 mL Heparin HiTrap column. Subsequently, fixed elution volumes were analyzed for the percentage of captured (a) HBsAg and (b) HBV GE compared to the totals present in the respective supernatant loaded. Error bars indicate five technical replicates [4]; (c) A single 5 mL Heparin HiTrap column was loaded with 275 mL of HBV-rich supernatant and eluted in 50 mL of elution buffer with different NaCl concentrations. Elution products were analyzed for protein concentration, HBsAg and HBV GE [4,31]. The dotted line indicates the suggested NaCl concentration for elution buffer (approx. 390 nM); (d) Similar to panel c, but showing the cumulative percentage of total for each readout; (e) Four 5 mL Heparin HiTrap columns were loaded with 550 mL of HBV-rich supernatant in parallel and afterwards eluted in a serial connection. The eluate was fractionated in 2 mL fractions and HBsAg was measured. The dotted lines show the start and end of suggested volume to collect (fraction 12–29 mL).