title: Validation of a biotherapeutic immunoaffinity-LC-MS/MS assay in monkey serum: 'plug-and-play' across seven molecules.
abstract: BACKGROUND
  Biotherapeutics development requires validated assays in biological matrices for pharmacokinetic assessment. Historically, ligand-binding assays have been the predominant platform available. Recently, alternative hybrid methods, combining ligand-binding analyte enrichment with LC-MS detection have emerged. Methodology & results: The validation of an immunoaffinity (IA)-LC-MS/MS method to quantify a monoclonal antibody biotherapeutic in cynomolgus monkey serum is described. This method includes immunoaffinity capture of the antibody in serum, followed by enzymatic digestion and detection of a framework peptide. Using similar method conditions, six additional biotherapeutic assays were readily validated in different nonhuman mammalian species, including mouse, rat and monkey.
  
  
  CONCLUSIONS
  The immunoaffinity-LC-MS/MS assay validation results across seven antibody therapeutics, using comparable conditions, illustrate the 'plug-and-play' nature of the IA-LC-MS/MS mAb framework peptide assay format.
authors.affiliation: Genentech, Inc., South San Francisco, California, USA.
authors.forename: Surinder
authors.initials: S
authors.lastname: Kaur
authors.affiliation: Genentech, Inc., South San Francisco, California, USA.
authors.forename: Luna
authors.initials: L
authors.lastname: Liu
authors.affiliation: PPD Laboratories, Richmond, Virginia, USA.
authors.forename: Diego F
authors.initials: DF
authors.lastname: Cortes
authors.affiliation: PPD Laboratories, Richmond, Virginia, USA.
authors.forename: Junlong
authors.initials: J
authors.lastname: Shao
authors.affiliation: PPD Laboratories, Richmond, Virginia, USA.
authors.forename: Rand
authors.initials: R
authors.lastname: Jenkins
authors.affiliation: PPD Laboratories, Richmond, Virginia, USA.
authors.forename: William R
authors.initials: WR
authors.lastname: Mylott
authors.affiliation: Genentech, Inc., South San Francisco, California, USA.
authors.forename: Keyang
authors.initials: K
authors.lastname: Xu
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood*; D061067:Antibodies, Monoclonal, Humanized / Q000097:blood; D002846:Chromatography, Affinity / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D008252:Macaca fascicularis / Q000097:blood; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized
keywords: LC–MS/MS; biotherapeutic; calibration standard; high performance LC; immunoaffinity; lower limit of quantification; percent coefficient of variation; quality control; standard deviation; upper limit of quantification; validation