Selective clearance of glycoforms of a complex glycoprotein pharmaceutical caused by terminal N-acetylglucosamine is similar in humans and cynomolgus monkeys.

摘要: To understand how the carbohydrate moieties of a recombinant glycoprotein affected its pharmacokinetic (PK) properties, glycan distribution was directly assessed from serial blood samples taken during PK studies in cynomolgus monkeys and humans. The protein studied an immunoadhesin (lenercept), containing Fc domain human immunoglobulin G (IgG-1) two copies extensively glycosylated extra cellular tumor necrosis factor receptor p55. recovered pure form using dual column, immunoaffinity-reversed-phase high-performance liquid chromatography method. glycans were released analyzed by matrix-assisted laser desorption ionization time flight mass spectrometry (MALDI-TOF MS). Alternatively, trypsin used to obtain glycopeptides, these MALDI-TOF. composition versus profiles show that not altered over 10 days circulation, consistent with their sequestration interior protein. However, changed dramatically first 24 h then remained relatively constant. Analysis acidic (derived exclusively domain) showed that, rapid initial phase clearance, carrying terminal N-acetylglucosamine (tGlcNAc) selectively cleared circulation. This phenomenon occurred similarly humans monkeys. Sialic acid content galactose only small changes. These data confirm correlation tGlcNAc half-life molecule, support hypothesis mannose (which can also bind tGlcNAc) causes variable clearance this molecule.