title: Large-scale implementation of sequential protein and peptide immunoaffinity enrichment LC/nanoLC-MS/MS for human β-nerve growth factor.
abstract: BACKGROUND
  A previously described immunoaffinity (IA)-LC-MS/MS assay for human β-nerve growth factor (β-NGF) was implemented to support large-scale sample testing for multiple clinical trials. Methodology & results: The procedure was modified to increase throughput by simultaneous preparation of two 96-well plates and LC duty-cycle reduction. Robustness of the LC method and nano-ESI was ensured during large-scale assay execution by closely monitoring and, if needed, replacing system components prior to failure. Following validation, the assay was used to analyze approximately 19,000 samples from multiple clinical studies over several years.
  
  
  CONCLUSIONS
  Routine implementation of the β-NGF IA-LC-MS/MS assay supported drug development programs. This optimized assay format now serves as a template for other clinical protein biomarker assays.
authors.affiliation: Q Squared Solutions BioSciences LLC, Ithaca, NY 14850, USA.
authors.forename: Gary A
authors.initials: GA
authors.lastname: Schultz
authors.affiliation: Q Squared Solutions BioSciences LLC, Ithaca, NY 14850, USA.
authors.forename: Kathlyn
authors.initials: K
authors.lastname: McCardle
authors.affiliation: Pharmacokinetics Dynamics & Metabolism, Pfizer, Inc., Andover, MA 01810, USA.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Neubert
mesh_terms: D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D006801:Humans; D036103:Nanotechnology / Q000592:standards; D020932:Nerve Growth Factor / Q000032:analysis* / Q000302:isolation & purification / Q000592:standards; D011786:Quality Control; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification / Q000592:standards; D053719:Tandem Mass Spectrometry / Q000592:standards
chemical_list: C502240:NGF protein, human; D011994:Recombinant Proteins; D020932:Nerve Growth Factor
keywords: LC–MS; antipeptide antibody column; biomarkers; extended SIL peptide; immunoaffinity; method development