title: Profiling the effects of process changes on residual host cell proteins in biotherapeutics by mass spectrometry.
abstract: An advanced liquid chromatography/mass spectrometry (MS) platform was used to identify and quantify residual Escherichia coli host cell proteins (HCPs) in the drug substance (DS) of several peptibodies (Pbs). Significantly different HCP impurity profiles were observed among different biotherapeutic Pbs as well as one Pb purified via multiple processes. The results can be rationally interpreted in terms of differences among the purification processes, and demonstrate the power of this technique to sensitively monitor both the quantity and composition of residual HCPs in DS, where these may represent a safety risk to patients. The breadth of information obtained using MS is compared to traditional multiproduct enzyme-linked immunosorbent assay (ELISA) values for total HCP in the same samples and shows that, in this case, the ELISA failed to detect multiple HCPs. The HCP composition of two upstream samples was also analyzed and used to demonstrate that HCPs that carry through purification processes to be detectable in DS are not always among those that are the most abundant upstream. Compared to ELISA, we demonstrate that MS can provide a more comprehensive, and accurate, characterization of DS HCPs, thereby facilitating process development as well as more rationally assessing potential safety risks posed by individual, identified HCPs.
authors.affiliation: Dept. of Process and Product Development, Amgen Inc., Thousand Oaks, CA, USA.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Schenauer
authors.forename: Gregory C
authors.initials: GC
authors.lastname: Flynn
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Goetze
mesh_terms: D001709:Biotechnology; D002853:Chromatography, Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D013058:Mass Spectrometry; D011506:Proteins / Q000302:isolation & purification*
chemical_list: D011506:Proteins
keywords: biotherapeutics; host cell proteins; mass spectrometry