Hydrolysis of Polysorbate 20 and PS80 with Putative
Phospholipase B-like 2 (PLBD2) and Formulated Antibody
To evaluate the effect of PLBD2 on PS20 and PS80 degradation, 16 µL of
200 mg/mL mAb-3 in 10 mM Histidine buffer pH 6.0 was mixed with 2 µL of
1% PS20 or 1% PS80, followed by adding 2 µL of 0.2 mg/mL PLBD2 and
incubating the sample at 45°C for 5 days. One aliquot (3 µL) of each
sample was diluted 25-fold by adding 72 µL 10 mM histidine, pH 6.0 and
used for LC-CAD analysis.
The hydrolysis of PS20 in formulated mAb was examined by mixing 18 µL of
75 mg/mL mAb (buffer exchanged to 10 mM Histidine, pH 6.0) with 2 µL of
1% PS20 followed by incubation at 45°C for 5 days. The hydrolysis of
PS80 in formulated mAb was examined by mixing 18 µL of 100 mg/mL mAb
(buffer exchanged to 10mM Histidine, pH 6.0) with 2 µL of 1% PS80
followed by incubation at 45°C for 5 days. One aliquot (3µL) of each
sample was diluted 25-fold by 10 mM Histidine, pH 6 and used for LC-CAD
analysis.