Figure 6

FGF-2 ligand blot binding of glycosidase-digested rhPln.D1. Pooled anionic exchange fractions of rhPln.198 or rhPln.247 from HEK 293 cell culture were subjected to digestion as designated; buffer only, heparinase I (Hep I), heparinase II (Hep II), heparinase III (Hep III), chondroitinase ABC (Chond. abc), all three heparinases plus chondroitinase ABC, or heparanase. Samples were subjected to SDS-PAGE and blotted to nitrocellulose. After blocking, the blot was then incubated with rhFGF-2 overnight, washed, then the bound rhFGF-2 was detected by anti-FGF-2 biotin-conjugates and streptavidin-alkaline phosphatase staining.