title: A universal surrogate peptide to enable LC-MS/MS bioanalysis of a diversity of human monoclonal antibody and human Fc-fusion protein drug candidates in pre-clinical animal studies.
abstract: For the development of human antibody Fc (fraction crystallizable) region-containing therapeutic protein candidates, which can be either monoclonal antibodies (mAbs) or pharmacologically active proteins/peptides fused to the Fc region of human Immunoglobulin G (IgG), reliable quantification of these proteins in animal pharmacokinetic study plasma samples is critical. LC-MS/MS has emerged as a promising assay platform for this purpose. LC-MS/MS assays used for bioanalysis of human antibody Fc region-containing therapeutic protein candidates frequently rely upon quantification of a 'signature' surrogate peptide whose sequence is unique to the protein analyte of interest. One drawback of the signature peptide approach is that a new LC-MS/MS assay must be developed for each new human Fc region-containing therapeutic protein. To address this issue, we propose an alternative 'universal surrogate peptide' approach for the quantification of human antibody Fc region-containing therapeutic protein candidates in plasma samples from all nonclinical species. A single surrogate tryptic peptide was identified in the Fc region of most human antibody Fc-containing therapeutic protein candidates. An LC-MS-MS method based upon this peptide was shown to be capable of supporting bioanalysis of a diversity of human Fc region-containing therapeutic protein candidates in plasma samples of all commonly used animal species.
authors.affiliation: Analytical Research and Development, Research and Development, Bristol-Myers Squibb, Princeton, NJ 08543-4000, USA.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Furlong
authors.forename: Zheng
authors.initials: Z
authors.lastname: Ouyang
authors.forename: Steven
authors.initials: S
authors.lastname: Wu
authors.forename: James
authors.initials: J
authors.lastname: Tamura
authors.forename: Timothy
authors.initials: T
authors.lastname: Olah
authors.forename: Adrienne
authors.initials: A
authors.lastname: Tymiak
authors.forename: Mohammed
authors.initials: M
authors.lastname: Jemal
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics; D001216:Asparagine / Q000737:chemistry / Q000378:metabolism; D001224:Aspartic Acid / Q000737:chemistry / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D003198:Computer Simulation; D004285:Dogs; D055808:Drug Discovery / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006168:Guinea Pigs; D006801:Humans; D006868:Hydrolysis; D007141:Immunoglobulin Fc Fragments / Q000097:blood / Q000737:chemistry* / Q000378:metabolism; D008252:Macaca fascicularis; D008253:Macaca mulatta; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000097:blood / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics; D011817:Rabbits; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods; D014357:Trypsin / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D007141:Immunoglobulin Fc Fragments; D010446:Peptide Fragments; D001224:Aspartic Acid; D001216:Asparagine; D014357:Trypsin