Erythropoietin is a therapeutic glycoprotein that stimulates red blood cell production. The quality, safety, and potency of recombinant erythropoietins are determined largely by their glycosylation. Small variations in cell culture conditions can significantly affect the glycosylation, and therefore the efficacy, of recombinant erythropoietins. Thus, detailed glycomic analyses are necessary to assess biotherapeutic quality. We have developed a platform for qualitative and quantitative glycomic analysis of recombinant erythropoietins. The platform was used to profile native N-glycans from three different type erythropoietin a common form of recombinant erythropoietin. Darbepoetin alfa was found to contain an abundance of large, multi-antennary N-glycans with high levels of sialylation, O-acetylation, and dehydration. Results were verified by independent orthogonal analysis with both MALDI-TOF and nano-LC/Q-TOF MS. This platform may be applied to quality control and batch analysis of not only recombinant erythropoietin but also other complex, glycosylated biotherapeutics and biosimilars.