Sequential protein and peptide immunoaffinity capture for mass spectrometry-based quantification of total human β-nerve growth factor

  • Neubert H
  • Muirhead D
  • Kabir M
 et al. 
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Abstract

Nerve growth factor (NGF) is a neurotrophin that is implicated in the modulation of pain perception. Tanezumab, a humanized monoclonal antibody (mAb) specific for NGF, is highly potent in sequestering NGF and has demonstrated efficacy for treatment of chronic pain in clinical trials. We describe a novel, sensitive immunoaffinity liquid chromatography−tandem mass spectrometry (LC−MS/MS) assay for quantitative determination of human serum NGF levels at baseline and after tanezumab treatment. The assay combines magnetic bead-based NGF immunoaffinity enrich- ment using a non-neutralizing polyclonal antibody followed by digestion and quantitation of a NGF-derived tryptic peptide via high-flow peptide immunoaffinity enrichment and nanoflow LC−MS/MS. Following validation, the assay was employed to measure total NGF concentrations in samples from clinical studies. The assay had a

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Authors

  • Hendrik Neubert

  • David Muirhead

  • Musarat Kabir

  • Chris Grace

  • Adriaan Cleton

  • Rosalin Arends

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