earticle

영어

Recombinant erythropoietin (EPO) is an important biotherapeutic used to treat anemia. However, the quality, safety, and potency of EPO are determined largely by its glycosylation, which is in turn strongly affected by the conditions of the cell culture in which it is produced. Qualitative and quantitative glycomic analyses of glycosylated biosimilars are important for assessing their quality as well as determining batch-to-batch variation during production. We studied overall profiling and comparative quantitation of native N-glycans from three batches of darbepoetin-alfa using MALDI-TOF MS and nano-LC/Q-TOF MS. After detergent removal, denaturation, and enzymatic N-glycan release, N-glycans were purified by graphitized carbon solid phase extraction (SPE). Darbepoetin-alfa was found to contain mostly tetraantennary, tetrasialylated glycans. Sialic acid modifications such as O-acetylation and dehydration were extensively observed, as were low abundances of polylactosamine-containing N-glycans. The correlation coefficients amongst the three batches were above 0.94. We observed similar results from both MALDI-TOF MS and nano-LC/Q-TOF MS. This platform was successfully applied to characterize the glycosylation of a complex biosimilar.