title: Combined use of immunoassay and two-dimensional liquid chromatography mass spectrometry for the detection and identification of metabolites from biotherapeutic pharmacokinetic samples.
abstract: Peptides and monoclonal antibodies have both emerged as important therapeutic modalities, but each has challenges which limit their use. Non-recombinant chemical conjugation of peptides onto antibodies has the potential to minimize or eliminate altogether many of these limitations. Once such approach, pioneered by CovX has created the possibility for rapid stoichiometric fusion of pharmacophores to a single antibody platform. These molecules, called CovX-Bodies, maintain both the pharmacologic properties of a given peptide and the pharmacokinetic properties of a monoclonal antibody. The result is a new class of molecules wherein each component contributes desirable traits. In this paper, we demonstrate the use of immunoassay and two-dimensional liquid chromatography mass spectrometry (2DLC/MS) in combination to investigate the antibody conjugates of Glucagon-like peptide-1 (GLP-1) and analogs for intact protein metabolite identification directly from mouse serum. The information gained from combining these approaches has helped guide and expedite the optimization of our drug product development efforts.
authors.affiliation: CovX, Pfizer Biotherapeutics Research and Development, United States. Robert.Murphy4@pfizer.com
authors.forename: Robert E
authors.initials: RE
authors.lastname: Murphy
authors.forename: Arvind G
authors.initials: AG
authors.lastname: Kinhikar
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Shields
authors.forename: Joselyn
authors.initials: J
authors.lastname: Del Rosario
authors.forename: Ryan
authors.initials: R
authors.lastname: Preston
authors.forename: Nancy
authors.initials: N
authors.lastname: Levin
authors.forename: Gary H
authors.initials: GH
authors.lastname: Ward
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000918:Antibody Specificity; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000077270:Exenatide; D052216:Glucagon-Like Peptide 1 / Q000097:blood*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000097:blood*; D014688:Venoms / Q000097:blood*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D010455:Peptides; D014688:Venoms; D052216:Glucagon-Like Peptide 1; D000077270:Exenatide