Analytical methods to quantitate and assess charge variants in recombinant monoclonal antibodies and their structural characterisation.
Mol weight of Mab is 150KDa. But on a 10% SDS Non-reducing PAGE it aligns above the 200KDa marker. But after reduction has two bands of approximately 50KDa and 25KDa. What could be the reason?
Glycosylation and marker variability are magnified in non-reducing native conditions - a more stringent technique will show ~155-165 kD for IgG isotypes.
2x55kD+2x25kD in reducing SDS gels.