Fig. 5
From: Secretion of Fc-amidated peptide fusion proteins by Chinese hamster ovary cells
Purification and mass spectroscopic analysis of Fc-fusion proteins. a After concentration using a Vivaflow flip flow filtration system and a Centricon, secreted Fc-fusion proteins were purified by binding to HiTrap Protein A cartridges; bound protein was eluted by application of low pH buffer and aliquots (E1-E4) were subjected to SDS-PAGE; purified Fc was analyzed at the same time. Comparison of glycan heterogeneity by intact mass analysis of Fc-AP-GLP1 (b) and Fc from manufacturing cell line (c). Peak labels are shorthand notation for glycosylation state. All molecules exhibited two bi-antennary core fucosylated glycans (BiF/BiF) with an additional Gn (n = 1-4) galactose and Sn (=1–2) sialic acid (N-acetylneuraminic acid) moieties distributed between the two glycans