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Table 2 Highlights of ECL antibody assay validation characteristics

From: Assessment of clinical immunogenicity of inotuzumab ozogamicin in patients with non-Hodgkin lymphoma and acute lymphoblastic leukemia

PC PC1: Rabbit Anti-InO
PC2: Rabbit Anti-CMC (Payload) Antibody
PCs in biological matrix (drug tolerance and confirmation assay) HPC 1:75 in 100% serum
LPC 1:750 in 100% serum
Intra-assay precision (%CV of endpoint log10 titer) 0.8%
Inter-assay precision (%CV of endpoint log10 titer) 1.2%
Sensitivity (affinity-purified polyclonal antibody) 215 ng/mL (in 100% serum)
Drug interference PC at high concentration was detected in the presence of 50 μg/mL; PC at low concentration was detected in the presence of 5.0 μg/mL
Matrix selectivity (recovery) with PCs All unspiked samples generated signal below the cut point showing negative result; 90% of ALL lots tested had recovery between 75% and 125% for both control antibodies
Stability (PC1 and PC2) 23-h ambient temperature
5 freeze/thaw cycles at − 70 °C and − 20 °C
Stability (biotinylated and ruthenylated InO freeze/thaw stability) 5 freeze/thaw cycles at − 70 °C
  1. %CV coefficient of variation, ALL acute lymphoblastic leukemia, CMC calicheamicin, ECL electrochemiluminescence, HPC high positive control, InO inotuzumab ozogamicin, LPC low positive control, PC positive control