Quantification of nivolumab in human plasma by LC-MS/HRMS and LC-MS/MS, comparison with ELISA

Nivolumab is a fully human immunoglobulin G4 used for the treatment of several advanced solid cancers as immune checkpoint inhibitors. There are some challenges quantification mAb in plasma because IgG present intrinsically complex biologic matrices and this determination must be based on reliable, selective, accurate analytical methods. This study described two validated methods carried out separate laboratories, one developed with triple quadrupole tandem mass spectrometry (LC-MS/MS) other high resolution an orbitrap system (LC-MS/HRMS). Both full-length stable isotope-labeled nivolumab-like (Arginine 13C6–15N4 Lysine 13C6–15N2) internal standard. The sample preparation was immunocapture, then trypsin digestion performed surrogate peptide quantified positive mode. Assays showed good linearity over range 5–100 ?g/mL 5–150 LC-MS/HRMS LC-MS/MS, respectively. limit set at 2 5 Acceptable accuracy (from - 13.6% to 3.0%) precision (within 20%) values were also obtained both LC-MS very different matrix effect linked use columns elution gradients. concentrations from 60 cancer outpatients compared home-made ELISA method. Bland–Altman analysis did not show any significant bias between three Passing–Bablock linear regression agreement better correlation