Fig 1.
Optimised protocols for PD-L1 IHC assays using the 22C3 antibody concentrate on the Dako ASL48 and VENTANA BenchMark ULTRA platforms.
PD-L1, programmed death ligand 1; IHC, immunohistochemistry; ASL48, Autostainer Link 48; DAB, 3,3’-diaminobenzidine tetrahydrochloride.
Fig 2.
PD-L1 staining patterns on NSCLC specimens using the 22C3 antibody concentrate on the Dako ASL48 platform (LDT) compared with the PD-L1 IHC 22C3 pharmDx kit on the Dako ASL48 platform (gold standard).
(A) The PD-L1 22C3 pharmDx kit on the Dako ASL48 platform; (B) optimised LDT (primary antibody dilution 1:50, 30-minute incubation); (C) LDT using primary antibody dilution 1:100, 30-minute incubation; (D) primary antibody dilution 1:200, 30-minute incubation; (E) primary antibody dilution 1:100, 60-minute incubation; (F) primary antibody dilution 1:100, 120-minute incubation. Original magnification 5×. Inserts, original magnification 40×. PD-L1, programmed death ligand 1; NSCLC, non–small cell lung cancer; ASL48, Autostainer Link 48; LDT, laboratory-developed test; IHC, immunohistochemistry.
Fig 3.
PD-L1 staining patterns on NSCLC specimens using the 22C3 antibody concentrate on the VENTANA BenchMark ULTRA platform (LDT) compared with the PD-L1 IHC 22C3 pharmDx kit on the Dako ASL48 platform (gold standard).
(A) The PD-L1 IHC 22C3 pharmDx kit on the Dako ASL48 platform; (B) optimised LDT (CC1 64 minutes, 22C3 primary antibody dilution 1:50, OptiView amplification 12 minutes); (C) CC1 32 minutes, primary antibody dilution 1:50, OptiView amplification 12 minutes; (D) CC1 64 minutes, primary antibody dilution 1:50, OptiView amplification 4 minutes; (E) CC1 32 minutes, primary antibody dilution 1:100, OptiView amplification 12 minutes; (F) CC1 64 minutes, primary antibody dilution 1:100, OptiView amplification 12 minutes. Original magnification 5×. Inserts, original magnification 40×. PD-L1, programmed death ligand 1; NSCLC, non–small cell lung cancer; ASL48, Autostainer Link 48; LDT, laboratory-developed test; IHC, immunohistochemistry.
Table 1.
PPA and NPA for each of the PD-L1 IHC assays using the 22C3 antibody concentrate on the Dako ASL48 and VENTANA BenchMark ULTRA platforms (LDTs) relative to the PD-L1 IHC 22C3 pharmDx kit on the Dako ASL48 platform (gold standard).
Table 2.
Intraclass correlation coefficient of TPS rating for each of the PD-L1 IHC assays using the 22C3 antibody concentrate on the Dako ASL48 and VENTANA BenchMark ULTRA platforms (LDTs) relative to the PD-L1 IHC 22C3 pharmDx kit on the Dako ASL48 platform (gold standard).