Depletion of PLBD2 from CHO-Derived Antibodies
PLBD2 depletion experiment was performed by using the Dynabeads antibody coupling kit as described by the manufacturer (Figure 5). Briefly, 5 mg of pre-washed magnetic Dynabeads were mixed with 100 µg anti-PLBD2 mAb in buffer C1. After addition of and buffer C2, the mixture was incubated overnight with gentle rocking at 4°C. Beads were washed by HB, LB and SB from the kit and then resuspend into 500 µL water. 50 µL resuspended anti-PLBD2 Dynabeads was added to each 10 mg mAb samples to a total volume of 500 µL respectively, followed by shaking at room temperature for 3 hours. After removing the beads, the supernatant was dried under SpeedVac and resuspended into water. Protein concentration of mAb was measured and adjusted to 75 mg/mL for incubation with 0.1% PS20 and 100 mg/mL for incubation with 0.1% PS80.