Posttranslational modification Trenonacog alfa Nonacog alfa Plasma-derived factor IX Disulfide bridging Eleven disulfide bridges Eleven disulfide bridges Eleven disulfide bridges [4 ] γ -Carboxylation≥11.7 mol γ -carboxyglutamic acid/mol factor IX 11.5 mol γ -carboxyglutamic acid/mol factor IX [4 ] 12 mol γ -carboxyglutamic acid/mol factor IX [5 ] β -Hydroxylation50% β -hydroxylated at Asp64 46% β -hydroxylated at Asp64 ~40% β -hydroxylated at Asp64 [4 ] Sulfation Tyr155: low degree of sulfation observed (5%) Tyr155: low degree of sulfation observed Tyr155: high degree of sulfation observed [4 ] Phosphorylation Ser158: low degree of phosphorylation observed (3%) Ser158: low degree of phosphorylation observed Ser158: ~90% phosphorylated [6 ] O-linked glycosylation EGF domain Ser53: -Glc Ser61: Neu5Ac-Hex-HexNAc-Fuc Ser53: -Glc [21 ] Ser61: Neu5Ac-Gal-GlcNAc-Fuc [21 ] Ser53: -Glc [7 ] Ser61: Neu5Ac-Gal-GlcNAc-Fuc [7 , 8 ] Activation peptide Low degree of site occupancy observed (13%) Low degree of site occupancy observed [4 ] Low degree of site occupancy observed [9 ] N-linked glycosylation N-glycans are complex, predominantly tri- and tetraantennary, and predominantly sialylated; 80% of N-glycans are either tetrasialylated or trisialylated N-glycans are complex, predominantly tri- and tetraantennary, and predominantly sialylated [4 ] N-glycans are complex, predominantly tri- and tetraantennary, and predominantly sialylated [10 ] Total sialylation 8.8 mol sialic acid/mol factor IX 6.5 mol sialic acid/mol factor IX [9 ] 8.8 mol sialic acid/mol factor IX [9 ]