Protein soluble aggregate assays
Soluble product aggregates were analyzed on a Waters UPLC system
(Waters, Acquity H-Class PLUS) with UV detection at 280 nm, using an
Acquity UPLC protein BEH SEC column (Waters, 125 Å, 1.7 μm, 4.6 mm×150
mm) and an Acquity UPLC protein BEH guard column (Waters, 4.6 mm × 30
mm). All samples were 0.2 μm filtered to remove potential large
particles prior to applying a total mass of 50 μg protein to the
SEC-UPLC system. The mobile phase (200 mM sodium phosphate, 150 mM
sodium chloride, pH 6.8) was run at a flow rate of 0.4 mL/min. Results
were analyzed using Waters Empower 3 chromatography data system software
(Waters, version 3).