Objective: To analyze residual CHO host cell protein (HCP) in human erythropoietin (hEPO) products by UHPLC-MS. Methods: The hEPO bulks and internal protein standard were denatured, reduced, alkylated and digested; Accucore Vanquish C18 column connected with UHPLC and Q Exactive Plus Hybrid Quadrupole-Orbitrap Mass Spectrometer were employed to separate and detect the samples; qualitation and quantitation of HCP were performed using HCP workflow in BioPharma Finder 4.1 software. Results: Ten HCPs were detected in all 4 batches of hEPO bulks, and the HCP amounts varied between different samples, ranging from 25×10-6 to approximately 1 000×10-6; the CV% were below 10% for the HCPs above 50×10-6. Conclusions: The method of UHPLC-MS could be used for qualitative and quantitative analysis of HCP in hEPO products expressed in CHO system, and the HCP types and amounts in different samples varied considerably.
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