Development of an automated PD-L1 immunohistochemistry (IHC) assay for non-small cell lung cancer

Therese Phillips, Pauline Simmons, Hector D Inzunza, John Cogswell, James Novotny Jr, Clive Taylor, Xiaoling Zhang, Therese Phillips, Pauline Simmons, Hector D Inzunza, John Cogswell, James Novotny Jr, Clive Taylor, Xiaoling Zhang

Abstract

Nivolumab, a fully human IgG4 programmed death 1 (PD-1) immune checkpoint inhibitor antibody, developed by Bristol-Myers Squibb Inc., has activity across non-small cell lung cancer (NSCLC) histologies and is Food and Drug Administration approved for treatment of metastatic squamous NSCLC with progression on or after platinum-based chemotherapy. PD-L1 has been investigated as a potential biomarker to predict clinical response to nivolumab in clinical settings. We report an automated PD-L1 immunohistochemistry (IHC) assay, which was developed to detect cell surface PD-L1 in formalin-fixed paraffin-embedded human tumor tissue specimens using Dako's Autostainer Link 48. The primary antibody for this assay is a rabbit monoclonal anti-human PD-L1 antibody, clone 28-8. The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines. The specificity of the PD-L1 IHC assay was further evaluated in a collection of 30 normal human tissues. The PD-L1 IHC assay was optimized for high sensitivity and precision in routine application. A pathology scoring and interpretation method specific to nivolumab clinical studies was adopted for the assay. The analytical performance of the assay was validated for application in the determination of PD-L1 status in human NSCLC specimens. The clinical application of the assay and scoring method was further validated in 3 Clinical Laboratory Improvement Amendments certified labs. The assay is currently being investigated in a variety of clinical studies for use as an in vitro diagnostic to select and stratify patients for treatment with the anti-PD-1 therapeutic antibody, nivolumab.

Conflict of interest statement

C.T. is a consultant to Dako North America contracted through Keck School of Medicine of the University of Southern California. The remaining authors declare no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Positive PD-L1 membrane staining in NSCLC tumor tissues illustrating intensity grades (top, ×20) and PD-L1 tumor scores (bottom, ×40). NSCLC indicates non–small cell lung cancer; PD-L1, programmed cell death 1 ligand 1.
FIGURE 2
FIGURE 2
Photomicrographs (×40) of HT-29 parental and PD-L1 overexpressing FFPE cell pellets stained by the PD-L1 IHC assay with and without antigen competition. FFPE indicates formalin-fixed paraffin-embedded; IHC, immunohistochemistry; NCR, negative control reagent; PD-L1, programmed cell death 1 ligand 1.
FIGURE 3
FIGURE 3
Photomicrographs (×40) of parental and PD-L1 knock-out (KO)-stable FFPE cell pellets stained by the PD-L1 IHC assay. FFPE indicates formalin-fixed paraffin-embedded; IHC, immunohistochemistry; NCR, negative control reagent; PD-L1, programmed cell death 1 ligand 1.

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Source: PubMed

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