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.