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

Phillips T, Simmons P, Inzunza HD, Cogswell J, Novotny J, Taylor C, Zhang X - Appl. Immunohistochem. Mol. Morphol. (2015)

Bottom Line: 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.The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines.

View Article: PubMed Central - PubMed

Affiliation: *Dako North America Inc., Carpinteria ‡Keck School of Medicine of the University of Southern California, Los Angeles, CA †Bristol-Myers Squibb Inc., Princeton, NJ.

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.

No MeSH data available.


Related in: MedlinePlus

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.
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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.

Mentions: The PD-L1 IHC assay specifically detects PD-L1 membrane protein expressed in tumor cells in FFPE blocks. HT-29 is a human colorectal tumor cell line that showed nondetectable PD-L1. Three stable HT-29 cell lines overexpressing full-length human PD-L1 were generated and showed differential expression of PD-L1 by FACS analysis (see the Materials and methods section). FFPE cell pellets of the parental and PD-L1 overexpressing cell lines were stained by the PD-L1 IHC assay. The level of PD-L1 detected by IHC was consistent with the PD-L1 level detected by FACS, and was abolished by the addition of PD-L1 antigen to the protein-rich antibody diluent (see the Materials and methods section, Fig. 2).


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

Phillips T, Simmons P, Inzunza HD, Cogswell J, Novotny J, Taylor C, Zhang X - Appl. Immunohistochem. Mol. Morphol. (2015)

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.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561627&req=5

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.
Mentions: The PD-L1 IHC assay specifically detects PD-L1 membrane protein expressed in tumor cells in FFPE blocks. HT-29 is a human colorectal tumor cell line that showed nondetectable PD-L1. Three stable HT-29 cell lines overexpressing full-length human PD-L1 were generated and showed differential expression of PD-L1 by FACS analysis (see the Materials and methods section). FFPE cell pellets of the parental and PD-L1 overexpressing cell lines were stained by the PD-L1 IHC assay. The level of PD-L1 detected by IHC was consistent with the PD-L1 level detected by FACS, and was abolished by the addition of PD-L1 antigen to the protein-rich antibody diluent (see the Materials and methods section, Fig. 2).

Bottom Line: 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.The specificity of 28-8 for PD-L1 was demonstrated by antigen competition and genetic deletion of PD-L1 in tumor cell lines.

View Article: PubMed Central - PubMed

Affiliation: *Dako North America Inc., Carpinteria ‡Keck School of Medicine of the University of Southern California, Los Angeles, CA †Bristol-Myers Squibb Inc., Princeton, NJ.

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.

No MeSH data available.


Related in: MedlinePlus