← Product Code [PLS](/productcode/PLS) · P150025S018 # PD-L1 IHC 28-8 pharmDx (P150025S018) _Agilent Technologies, Inc. · PLS · Jul 1, 2026 · APPR_ **Canonical URL:** https://fda-staging.innolitics.com/device/P150025S018 ## Device Facts - **Applicant:** Agilent Technologies, Inc. - **Product Code:** [PLS](/productcode/PLS.md) - **Decision Date:** Jul 1, 2026 - **Decision:** APPR - **Device Class:** Class 3 ## Indications for Use For in vitro diagnostic use. PD-L1 IHC 28-8 pharmDx is a qualitative immunohistochemical assay using Monoclonal Rabbit Anti-PD-L1, Clone 28-8 intended for use in the detection of PD-L1 protein in formalin-fixed, paraffin-embedded (FFPE) non-small cell lung cancer (NSCLC), squamous cell carcinoma of the head and neck (SCCHN), urothelial carcinoma (UC), gastric adenocarcinoma, gastroesophageal junction (GEJ) adenocarcinoma, and esophageal carcinoma tissues using EnVision FLEX visualization system on Autostainer Link 48. PD-L1 protein expression in NSCLC, SCCHN, and UC is determined by the percentage of evaluable tumor cells exhibiting partial or complete membrane staining at any intensity. PD-L1 protein expression in esophageal squamous cell carcinoma (ESCC), gastric adenocarcinoma, GEJ adenocarcinoma, and esophageal adenocarcinoma is determined by using Combined Positive Score (CPS), which is the number of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) divided by the total number of viable tumor cells, multiplied by 100. ## Device Story IHC assay using rabbit monoclonal anti-PD-L1 (clone 28-8) on FFPE tissue sections; utilizes EnVision FLEX visualization system on Dako Autostainer Link 48; detects PD-L1 protein expression via brown (DAB) reaction product. Pathologist evaluates staining using light microscopy; calculates Combined Positive Score (CPS) for ESCC, gastric, GEJ, and esophageal adenocarcinoma; evaluates tumor cell percentage for NSCLC, SCCHN, and UC. Output informs eligibility for nivolumab-based therapies. Used in clinical laboratories; assists in identifying patients likely to benefit from immunotherapy; mitigates risk of unnecessary toxicity or missed treatment opportunities. ## Clinical Evidence Clinical performance established via CHECKMATE-648 (ESCC) and CHECKMATE-649 (gastric/GEJ/esophageal adenocarcinoma) trials. CHECKMATE-648 (N=970) demonstrated improved OS for patients with CPS ≥ 1 treated with nivolumab-based regimens vs chemotherapy. CHECKMATE-649 (N=1581) demonstrated statistically significant OS improvement in patients with CPS ≥ 1 treated with nivolumab plus chemotherapy vs chemotherapy alone. Analytical validation included repeatability, reproducibility (inter-site/inter-observer), and robustness studies, confirming consistent performance across the CPS ≥ 1 range. ## Technological Characteristics IHC assay; rabbit monoclonal anti-PD-L1 (clone 28-8); EnVision FLEX visualization system (HRP-labeled dextran polymer); DAB+ chromogen; PT Link module for target retrieval (pH 6.1). Automated staining on Dako Autostainer Link 48. FFPE tissue input; 4-5 μm sections. Software: DakoLink v4.3.1. No electronic connectivity/cloud processing described. ## Regulatory Identification The programmed death-ligand 1 (PD-L1) antibody is a qualitative immunohistochemical antibody intended to identify PD-L1 protein expression in human clinical tissue specimens. The PD-L1 antibody is indicated as an aid in identifying patients eligible for treatment with specific FDA approved therapeutic drugs or to assess PD-L1 expression level in patients who may respond particularly well to specific FDA approved therapeutic drugs. ## Reference Devices - CHECKMATE-648 (CA209648) - CHECKMATE-649 (CA209649) - CHECKMATE-67T ## Submission Summary (Full Text) > This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com. > > Innolitics is a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices, including [a PMA](https://innolitics.com/services/regulatory/), [a 510(k)](https://innolitics.com/services/510ks/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/). {0} # SUMMARY OF SAFETY AND EFFECTIVENESS DATA (SSED) ## I. GENERAL INFORMATION Device Generic Name: In vitro diagnostic immunohistochemistry (IHC) for detection of PD-L1 protein in formalin-fixed, paraffin embedded (FFPE) human tissue sections Device Trade Name: PD-L1 IHC 28-8 pharmDx Device Procode: PLS Applicant's Name and Address: Agilent Technologies, Inc. 5301 Stevens Creek Blvd. Santa Clara, CA 95051 Date(s) of Panel Recommendation: None Premarket Approval Application (PMA) Number: P150025/S018 Date of FDA Notice of Approval: July 1, 2026 The original PMA (P150025) for PD-L1 IHC 28-8 pharmDx was approved on October 9, 2015, for the detection of PD-L1 protein in formalin-fixed, paraffin-embedded (FFPE) non-squamous non-small cell lung cancer (nsNSCLC) tissue using EnVision FLEX visualization system on Autostainer Link 48. A supplemental PMA (P150025/S003) for PD-L1 IHC 28-8 pharmDx indicated for squamous cell carcinoma of the head and neck (SCCHN) and urothelial carcinoma (UC) was approved on September 15, 2017. Subsequently, the indication for use was expanded to include detection of PD-L1 protein in NSCLC (non-squamous NSCLC and squamous NSCLC) patients for treatment with OPDIVO (nivolumab) in combination with YERVOY (ipilimumab) on May 15, 2020. The Summary of Safety and Effectiveness Data (SSEDs) for these indications are available on the CDRH website and are incorporated by reference here. The current supplement was submitted to expand the companion diagnostic (CDx) indication for PD-L1 IHC 28-8 pharmDx to include detection of PD-L1 protein in patients with esophageal squamous cell carcinoma (ESCC), and gastric, gastroesophageal junction (GEJ), and esophageal adenocarcinoma for treatment with OPDIVO (nivolumab) or OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy). ## II. INDICATIONS FOR USE For in vitro diagnostic use. PD-L1 IHC 28-8 pharmDx is a qualitative immunohistochemical assay using Monoclonal Rabbit Anti-PD-L1, Clone 28-8 intended for use in the detection of PD-L1 protein in formalin-fixed, ---PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 1 of 41 {1} paraffin-embedded (FFPE) non-small cell lung cancer (NSCLC), squamous cell carcinoma of the head and neck (SCCHN), urothelial carcinoma (UC), gastric adenocarcinoma, gastroesophageal junction (GEJ) adenocarcinoma, and esophageal carcinoma tissues using EnVision FLEX visualization system on Autostainer Link 48. PD-L1 protein expression in NSCLC, SCCHN, and UC is determined by the percentage of evaluable tumor cells exhibiting partial or complete membrane staining at any intensity. PD-L1 protein expression in esophageal squamous cell carcinoma (ESCC), gastric adenocarcinoma, GEJ adenocarcinoma, and esophageal adenocarcinoma is determined by using Combined Positive Score (CPS), which is the number of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) divided by the total number of viable tumor cells, multiplied by 100. Companion Diagnostic Indications | Tumor Indication | PD-L1 Expression Level | Therapy | | --- | --- | --- | | NSCLC | % Tumor Cell Expression ≥ 1 | OPDIVO® (nivolumab) | | ESCC | CPS ≥ 1 | OPDIVO® (nivolumab) or OPDIVO QVANTIG® (nivolumab and hyaluronidase-nvhy) | | Gastric, GEJ, or Esophageal Adenocarcinoma | CPS ≥ 1 | | When used in accordance with approved therapeutic labeling: PD-L1 expression (≥1% or ≥5% or ≥10% tumor cell expression), as detected by PD-L1 IHC 28-8 pharmDx in non-squamous NSCLC (nsNSCLC) may be associated with enhanced survival from OPDIVO®. PD-L1 expression (≥1% tumor cell expression), as detected by PD-L1 IHC 28-8 pharmDx in SCCHN may be associated with enhanced survival from OPDIVO®. PD-L1 expression (≥1% tumor cell expression), as detected by PD-L1 IHC 28-8 pharmDx in UC may be associated with enhanced response rate and enhanced disease-free survival from OPDIVO®. See the OPDIVO® and OPDIVO QVANTIG® product labels for specific clinical circumstances guiding PD-L1 testing. ### III. CONTRAINDICATIONS There are no known contraindications for use of this test. ### IV. WARNINGS AND PRECAUTIONS The warnings and precautions can be found in the PD-L1 IHC 28-8 pharmDx labeling. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 2 of 41 {2} # V. DEVICE DESCRIPTION PD-L1 IHC 28-8 pharmDx contains optimized reagents required to complete an immunohistochemical (IHC) staining procedure for formalin-fixed and paraffin-embedded (FFPE) specimens using the EnVision FLEX visualization system and the Dako Autostainer Link 48 automated staining system with DakoLink software. The principal component of the kit is the rabbit monoclonal anti-PD-L1 clone 28-8 antibody that binds to PD-L1 protein expressed on FFPE tissue. PD-L1 IHC 28-8 pharmDx includes reagents sufficient to perform 50 tests in up to 15 individual runs and instructions for use (IFU). A total of 28 vials containing reagents including the EnVision FLEX visualization system and 15 cell line control slides are provided in each kit. See Table 1 below. Table 1. Overview of PD-L1 IHC 28-8 pharmDx Components | Component | Description | Quantity x Volume | | --- | --- | --- | | Peroxidase-Blocking Reagent | Buffered solution containing hydrogen peroxide, detergent and 0.015 mol/L sodium azide. | 1 x 34.5 mL | | Primary Antibody: Monoclonal Rabbit anti-PD-L1, Clone 28-8 | Monoclonal rabbit anti-PD-L1 in a buffered solution, containing stabilizing protein, and 0.015 mol/L sodium azide. | 1 x 19.5 mL | | Negative Control Reagent (NCR) | Monoclonal rabbit control IgG antibody in a buffered solution, containing stabilizing protein, and 0.015 mol/L sodium azide. | 1 x 15 mL | | Linker, anti-Rabbit | Mouse secondary antibody against rabbit immunoglobulins in a buffered solution containing stabilizing protein and 0.015 mol/L sodium azide. | 1 x 34.5 mL | | Visualization Reagent-Horseradish Peroxidase (HRP) | Dextran coupled with peroxidase molecules and goat secondary antibody molecules against rabbit and mouse immunoglobulins in a buffered solution containing stabilizing protein and an antimicrobial agent. | 1 x 34.5 mL | | DAB+ Substrate Buffer | Buffered solution containing hydrogen peroxide and an antimicrobial agent. | 15 x 7.2 mL | | DAB+ Chromogen | 3,3'-diaminobenzidine tetrahydrochloride in an organic solvent. | 1 x 5 mL | | DAB Enhancer | Cupric sulfate in water. | 1 x 34.5 mL | | EnVision FLEX Target Retrieval Solution, Low pH, 50x | Buffered solution, pH 6.1, containing detergent and an antimicrobial agent (EnVision FLEX Target Retrieval Solution, Low pH, 50X). | 6 x 30 mL | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 3 of 41 {3} | Component | Description | Quantity x Volume | | --- | --- | --- | | PD-L1 IHC 28-8 pharmDx Control Slides | Each slide contains sections of two pelleted, formalin-fixed, paraffin-embedded cell lines: NCI-H226 with positive PD-L1 protein expression (originating from human lung squamous cell carcinoma with positive PD-L1 protein expression) and MCF-7 with negative PD-L1 protein expression (originating from human breast adenocarcinoma with negative PD-L1 protein expression). | 3 x 5 slides (15 slides) | ## Device Instrumentation and Software PD-L1 IHC 28-8 pharmDx assay is performed on the Dako Autostainer Link 48 automated staining system using the Dako Link software (version 4.3.1). The Autostainer system is designed to mimic the staining steps performed manually by a lab technician. The PD-L1 IHC 28-8 pharmDx protocol is assay specific. The DakoLink software has been designed to recognize and group PD-L1 IHC 28-8 pharmDx reagents, requiring that all system reagents are used together. Deparaffinization, rehydration, and target retrieval (3-in-1) procedures are performed in the Dako PT Link Pre-treatment module. ## Specimen Preparation Only FFPE tissues are suitable for use. Recommended handling and processing conditions are as follows: < 30 minutes ischemia time prior to immersion in fixative, and 24-48 hours fixation time in 10% neutral buffered formalin. Alternative fixative methods have not been validated and may give erroneous results. The use of PD-L1 IHC 28-8 pharmDx on decalcified tissues has not been validated and is not recommended. Specimens should be blocked into a thickness of 3 or 4 mm, fixed in 10% neutral buffered formalin, and dehydrated and cleared in a series of alcohols and xylene, followed by infiltration with melted paraffin. The paraffin temperature should not exceed 60 °C. Tissue specimens should be cut into sections of 4-5 μm. After sectioning, tissues should be mounted on Fisherbrand Superfrost Plus charged slides or FLEX IHC Microscope Slides (Code K8020) and then placed in a 58 ± 2 °C oven for 1 hour. To preserve antigenicity, tissue sections, once mounted on slides, should be stored in the dark at 2-8°C, or room temperature up to 25 °C. Slide storage and handling conditions should not exceed 25 °C at any point post-mounting to ensure tissue integrity and antigenicity. Gastric, GEJ, and esophageal carcinoma cut sections must be stained within 4 months when stored at 2-8°C or 25 °C. ## Test Controls Run controls are included in each staining run to establish the validity of the test results. The following controls must be run with the assay: 1) Control cell line slides provided as part of the kit should be used to verify the staining procedure. One Control Slide should be stained with the primary antibody to PD-L1 in each staining run. The evaluation of the Control Slide cell lines supplied in the kit indicates the PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 4 of 41 {4} validity of the staining run. The Control Slides should not be used as an aid in interpretation of patient results. 2) Positive run control tissue of the same tumor indication as the patient specimen are to be provided by the end-user laboratory. It is recommended that tissue that stains a weak to moderate intensity for PD-L1 is used to monitor for all aspects of pre-analytical variables such as fixation, processing, and sectioning. Negative control tissue of the same tumor indication as the patient specimen is required to detect unintended antibody cross reactivity to tissue and is expected to be negative for PD-L1 expression. Tissue features located in patient tissue or positive control tissue that is not expected to show PD-L1 expression may be used as negative control for staining. 3) The kit includes a Negative Control Reagent that is used in parallel with the PD-L1 Clone 28-8 primary antibody on patient tissue. The matched negative control reagent aids the reader in differentiating specific staining at the antigen site from tissue-specific background staining that occurs from reaction with detection chemistry and not the anti PD-L1 primary antibody. Additional information about the use of controls is available in the product labeling. ### Principle of Operation PD-L1 IHC 28-8 pharmDx contains optimized reagents and protocol required to complete an IHC staining procedure of FFPE specimens using Autostainer Link 48 and PT Link Pre-treatment Module. Patient FFPE tissue specimens are subject to deparaffinization, rehydration, and target retrieval in the Target Retrieval Solution Low pH (3-in-1) for 20 minutes at 97 °C in the PT Link instrument to expose the PD-L1 antigen if present on the tissue. The slides are then loaded onto the Autostainer Link 48 automated stainer and incubated with the primary monoclonal antibody to PD-L1 (Clone 28-8) or the Negative Control Reagent (NCR). This step enables binding of the primary antibody to tumor tissue section when PD-L1 antigen is expressed. The slides are then incubated with an anti-Rabbit linker antibody, specific to Fc region of the primary antibody. Following this, the slides are incubated with a ready-to-use visualization reagent consisting of secondary antibody molecules and horseradish peroxidase molecules coupled to a dextran polymer backbone. The enzymatic conversion of the subsequently added DAB+ chromogen results in precipitation of a visible reaction product at the antigen sites. The color of the chromogenic reaction is modified by a chromogen enhancement reagent, DAB Enhancer. The specimens are then counterstained with hematoxylin and cover slipped and observed under a microscope to visually determine if the PD-L1 protein is expressed in patient tumor tissue. ### Staining Protocol: The staining protocol for PD-L1 IHC 28-8 pharmDx on the Dako Autostainer Link 48 is as follows: - Rinse in buffer - Peroxidase-Blocking Reagent (2 drop zones x 150 μL): 5 minutes - Rinse in buffer PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 5 of 41 {5} - Monoclonal Rabbit anti-PD-L1 (or Negative Control Reagent) (2 drop zones x 150 μL): 30 minutes - Rinse in buffer - Linker Reagent (2 drop zones x 150 μL): 30 minutes - Rinse in buffer - Visualization Reagent-HRP (2 drop zones x 150 μL): 30 minutes - Rinse in buffer: 5 minutes - DAB+ solution (2 drop zones x 150 μL): 2 x 5 minutes - Rinse in buffer - DAB Enhancer (2 drop zones x 150 μL): 5 minutes - Rinse in buffer - Hematoxylin (2 drop zones x 150 μL): 7 minutes - Rinse in deionized water - Rinse in buffer: 5 minutes - Rinse in deionized water ## Interpretation of PD-L1 Staining for ESCC, and Gastric, GEJ, and Esophageal Adenocarcinoma Specimens Interpretation of PD-L1 IHC 28-8 pharmDx stained slides should be performed by a pathologist using a light microscope. PD-L1 staining is indicated with a brown (DAB) reaction product. Evaluation of PD-L1 expression by CPS is the number of PD-L1 staining cells (tumor cells, lymphocytes, macrophages) divided by the total number of viable tumor cells, multiplied by 100. Distinction of viable tumor cells, lymphocytes, and macrophages is essential for accurate denominator estimation. Although the result of the calculation can exceed 100, the maximum score is defined as CPS 100. CPS is defined as follows: $$\text{CPS} = \frac{\# \text{ PD-L1 staining cells (tumor cells, lymphocytes, macrophages)}}{\text{Total \# of viable tumor cells}} \times 100$$ For each staining run, slides should be examined in the order recommended in the labeling to determine the validity of the staining run and enable assessment of the staining of the sample tissue. Patient specimens stained with PD-L1 and the NCR from PD-L1 IHC 28-8 pharmDx should be examined when evaluating PD-L1 expression. Specimens stained with NCR must have 0 specific staining and ≤ 1+ nonspecific staining intensity. For evaluation of the immunohistochemical staining, an objective of 10-20x magnification is appropriate. For determination of PD-L1 expression, an objective of 20x magnification is required. By definition, PD-L1 staining cells are: - Tumor cells with convincing partial or complete linear membrane staining (at any intensity) that is perceived distinct from cytoplasmic staining and - Lymphocytes and macrophages (mononuclear inflammatory cells, MICs) within the tumor nests and/or adjacent supporting stroma with convincing membrane and/or PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 6 of 41 {6} cytoplasmic staining (at any intensity). MICs must be directly associated with the response against the tumor. The entire specimen must be evaluated. All viable tumor cells on the entire PD-L1-stained patient slide must be evaluated and included in the PD-L1 scoring assessment together with tumor associated PD-L1 positive lymphocytes and macrophages. A minimum of 100 viable tumor cells must be present in the PD-L1 stained slide (biopsy and resection) for the specimen to be considered adequate for evaluation. If patient specimens include more than one biopsy on a slide, all tissues on the slide need to be evaluated to generate a single CPS for determining the PD-L1 expression level. Each biopsy should not be reported independently. For specimens with less than 100 viable tumor cells, tissue from a deeper level of the block or potentially another block, could present sufficient tumor cells for PD-L1 evaluation. This assay was validated for invasive ESCC, gastric, GEJ, and esophageal adenocarcinoma tissue samples and not for lesions with foci of dysplasia or carcinoma in situ. H&E-stained slides should accompany each PD-L1 stained sample to allow proper assessment of invasive carcinoma, carcinoma in situ, and adjacent normal epithelium. The CPS denominator includes all viable invasive tumor cells (PD-L1 staining and non-staining). Dysplasia, carcinoma in situ, and all other cells are excluded. Table 2 provides details about which tissue elements are included in and excluded from the CPS numerator in ESCC, and gastric, GEJ, and esophageal adenocarcinoma specimens. Table 2. CPS Numerator Inclusion/Exclusion Criteria for ESCC, and Gastric, GEJ and Esophageal Adenocarcinoma | Tissue Elements | Included in the Numerator | Excluded from the Numerator | | --- | --- | --- | | Tumor cells | • Convincing partial linear or complete circumferential membrane staining (at any intensity) of viable invasive and metastatic esophageal carcinoma, gastroesophageal junction carcinoma or gastric carcinoma | • Non-staining tumor cells • Tumor cells with only cytoplasmic staining • Dysplasia • In situ carcinoma | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 7 of 41 {7} | Tissue Elements | Included in the Numerator | Excluded from the Numerator | | --- | --- | --- | | **Immune cells** | - Membrane and/or cytoplasmic* staining (at any intensity) of mononuclear inflammatory cells (MICs) within tumor nests and adjacent supporting stroma†: - Lymphocytes (including lymphocyte aggregates) - Macrophages‡ - Only MICs within 20X field and directly associated with the response to the tumor are scored | - Non-staining MICs - MICs (including lymphoid aggregates) not associated with the tumor - MICs associated with benign structures - Neutrophils, eosinophils and plasma cells | | **Other Cells** | - Not included | - Normal cells - Stromal cells (including fibroblasts) - Necrotic cells and/or cellular debris | * In MICs, membrane and cytoplasmic staining are often indistinguishable due to high nuclear to cytoplasmic ratio. Therefore, membrane and/or cytoplasmic staining of MICs is included in the CPS numerator † Adjacent MICs are defined as being within the same 20x field as the tumor. However, MICs that are NOT directly associated with the response to the tumor should be excluded ‡ Macrophages and histiocytes are considered the same cells The ESCC, and gastric, GEJ, and esophageal adenocarcinoma specimens should be considered to have PD-L1 expression if CPS ≥ 1. To accurately assess PD-L1 expression status, challenging borderline cases (slides) with scores near the CPS ≥ 1 cutoff (CPS > 0 and < 10) should be reviewed by an additional pathologist and the reported results should be based on a consensus score. ### VI. ALTERNATIVE PRACTICES AND PROCEDURES There are no other FDA-cleared or approved alternatives for the assessment of PD-L1 expression levels in FFPE human ESCC, and gastric, GEJ, and esophageal adenocarcinoma specimens to guide treatment with OPDIVO (nivolumab) or OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy). ### VII. MARKETING HISTORY PD-L1 IHC 28-8 pharmDx, as approved under P150025 in the US, is currently marketed in Argentina, Australia, Austria, Bahrain, Belgium, Brazil, Bulgaria, Canada, Chile, China, Colombia, Costa Rica, Croatia, Cyprus, Czech Republic, Denmark, Ecuador, Egypt, Estonia, PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 8 of 41 {8} Finland, France, Germany, Greece, Hong Kong, Hungary, Iceland, India, Ireland, Israel, Italy, Japan, Jordan, South Korea, Kosovo, Kuwait, Latvia, Lebanon, Liechtenstein, Lithuania, Luxembourg, Macau, , Malaysia, Malta, Mexico, , Netherlands, New Zealand, Norway, Oman, Paraguay, Peru, Poland, Portugal, Qatar, Romania, Saudi Arabia, Serbia, Singapore, Slovakia, Slovenia, Spain, Sweden, Switzerland, Taiwan, Thailand, Turkey, UAE, United States, United Kingdom, Uruguay. ### VIII. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH Failure of the device to perform as expected, or failure to correctly interpret test results, may lead to incorrect PD-L1 results and subsequently lead to improper treatment decisions for patients being considered for OPDIVO (nivolumab) or OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy) therapy. Patients with a false negative result may be excluded from potentially beneficial treatment with OPDIVO or OPDIVO QVANTIG. Patients with a false-positive result may receive OPDIVO or OPDIVO QVANTIG without an expectation of clinical benefit, unnecessarily exposing them to potential toxicity. Additionally, delayed test results may postpone the initiation of treatment. For the specific adverse events that occurred in the clinical studies, refer to the OPDIVO (nivolumab) and OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy) FDA-approved package inserts, which are available at Drugs@FDA. ### IX. SUMMARY OF NON-CLINICAL STUDIES #### A. Laboratory Studies Non-clinical studies were performed using PD-L1 IHC 28-8 pharmDx to establish analytical performance of the device. The scoring algorithm used in these studies resulted in a PD-L1 positive or negative output with an underlying score of CPS 0-100. Antibody characterization studies for clone 28-8, including specificity and tour of body/tour of tumor, control cell line validation, kit stability, and preanalytical variables were submitted and reviewed in the original PMA (P150025) for this device. Study designs and results are available in the Summary of Safety and Effectiveness Data (SSED) for the original PMA, https://www.accessdata.fda.gov/cdrh_docs/pdf15/P150025B.pdf. Results from studies performed to support the ESCC, and gastric, GEJ, and esophageal adenocarcinoma indications are summarized in the sections below. #### 1. Analytical Specificity Assessment of analytical specificity of the monoclonal rabbit anti-human PD-L1 clone 28-8 antibody was reviewed under P150025 and included Western blot and immunoreactivity in human tissues, both normal and tumor. Refer to the Summary of Safety and Effectiveness Data for P150025. ---PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 9 of 41 {9} ## 2. Analytical Sensitivity Analytical sensitivity of PD-L1 IHC 28-8 pharmDx was evaluated using 1226 unique FFPE specimens from cases of gastric cancer, GEJ cancer, and esophageal cancer (439 gastric, 375 GEJ, and 412 esophageal carcinoma), staged I to IV, using manufactured production lots. Assessment of PD-L1 expression demonstrated staining across a range of 0-100% positive tumor cells and staining intensity of 0-3. The number of positive cases at the CPS ≥ 1 expression level was 267 (60.8%), 219 (58.4%), and 295 (71.6%), respectively. ## 3. Repeatability The objective of this study was to demonstrate that PD-L1 IHC 28-8 pharmDx would produce consistent staining in normal day-to-day testing. Inter-instrument, inter-operator, inter-day, and intra-lot analyses were performed. All gastric, GEJ, and esophageal carcinoma specimens were stained, then blinded and randomized prior to evaluation of PD-L1 expression status. Specimens were determined to be PD-L1-positive if CPS ≥ 1 or PD-L1-negative if CPS < 1. Statistical analysis was conducted on the dichotomized data. Negative percent agreement (NPA), positive percent agreement (PPA), and overall agreement (OA) with two-sided 95% bootstrap confidence intervals were calculated for each of the study endpoints. A summary of the methods and results of the repeatability study is presented in Table 3. Per-specimen repeatability results are presented in Table 4. **Table 3. Summary of Precision (one site) for CPS ≥ 1** | Precision Endpoint | Study Design | % Agreement (95% CI) | | --- | --- | --- | | Combined Precision (inter-operator, inter-instrument, inter-day, and inter-lot as combined variables) | Each of 60 specimens spanning the full range of PD-L1 IHC expression (27 PD-L1-negative and 33 PD-L1-positive, comprising gastric, GEJ, and esophageal carcinoma) was tested under the following conditions: three operators, three instruments, three days, and three assay lots. A total of 180 independent comparisons to consensus (majority call) were performed. | NPA 96.3% (92.6, 100.0) PPA 98.0% (94.9, 100.0) OA 97.2% (94.9, 99.4) | NPA= Negative Percent Agreement; PPA= Positive Percent Agreement; OA=Overall Percent Agreement; CPS=Combined Positive Score To accurately assess PD-L1 expression status, challenging borderline cases (slides) with scores near the CPS ≥ 1 cutoff (CPS > 0 and < 10) should be reviewed by an additional pathologist and the reported results should be based on a consensus score. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 10 of 41 {10} **Table 4. Combined Precision Results per Specimen (inter-instrument/operator/day/lot)** | Specimen | Subtype | Number of Results | Consensus Result* | Median (CPS) | Range (CPS) | Between Day (SD) | Percent Positive Results** (n/N) | Percent Agreement with Consensus (n/N) | 95% CI*** | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 1 | GC | 3 | Negative | 0 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 2 | ESCC | 3 | Negative | 0 | 0, 0 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 3 | ESCC | 3 | Negative | 0 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 4 | GC | 3 | Negative | 0 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 5 | ESCC | 3 | Negative | 0 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 6 | GC | 3 | Negative | 0 | 0, 0 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 7 | GC | 3 | Negative | 0 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 8 | EAC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 9 | EAC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 10 | EAC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 11 | ESCC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 12 | GEJ | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 13 | EAC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 14 | GC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 15 | GC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 16 | GC | 3 | Negative | <1 | 0, <1 | 0.289 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 17 | ESCC | 3 | Negative | <1 | 0, 3 | 1.607 | 33.3% (1/3) | 66.7% (2/3) | 20.8-93.9% | | 18 | GEJ | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 19 | GEJ | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 20 | ESCC | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 21 | GC | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 22 | ESCC | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 23 | ESCC | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 24 | ESCC | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 25 | ESCC | 3 | Negative | <1 | <1, <1 | 0.000 | 0% (0/3) | 100% (3/3) | 43.9-100% | | 26 | GC | 3 | Negative | <1 | <1, 1 | 0.289 | 33.3% (1/3) | 66.7% (2/3) | 20.8-93.9% | | 27 | ESCC | 3 | Negative | <1 | <1, 5 | 2.598 | 33.3% (1/3) | 66.7% (2/3) | 20.8-93.9% | | 28 | EAC | 3 | Positive | 1 | <1, 1 | 0.289 | 66.7% (2/3) | 66.7% (2/3) | 20.8-93.9% | | 29 | GC | 3 | Positive | 1 | <1, 1 | 0.289 | 66.7% (2/3) | 66.7% (2/3) | 20.8-93.9% | | 30 | GC | 3 | Positive | 1 | 1, 5 | 2.309 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 31 | GC | 2 | Positive | 3 | 1, 5 | 2.828 | 100% (2/2) | 100% (2/2) | 34.2-100% | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 11 of 41 {11} | Specimen | Subtype | Number of Results | Consensus Result* | Median (CPS) | Range (CPS) | Between Day (SD) | Percent Positive Results** (n/N) | Percent Agreement with Consensus (n/N) | 95% CI*** | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 32 | EAC | 3 | Positive | 5 | 1, 10 | 4.509 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 33 | GC | 3 | Positive | 5 | 1, 10 | 4.509 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 34 | GC | 3 | Positive | 20 | 1, 30 | 14.731 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 35 | GC | 3 | Positive | 20 | 2, 30 | 14.189 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 36 | EAC | 3 | Positive | 50 | 2, 60 | 31.005 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 37 | ESCC | 3 | Positive | 5 | 5, 10 | 2.887 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 38 | GC | 3 | Positive | 5 | 5, 5 | 0.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 39 | ESCC | 3 | Positive | 5 | 5, 5 | 0.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 40 | GC | 3 | Positive | 10 | 5, 15 | 5.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 41 | GEJ | 3 | Positive | 10 | 10, 10 | 0.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 42 | GC | 3 | Positive | 10 | 10, 20 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 43 | ESCC | 3 | Positive | 30 | 10, 40 | 15.275 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 44 | ESCC | 3 | Positive | 50 | 10, 70 | 30.551 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 45 | EAC | 3 | Positive | 20 | 20, 40 | 11.547 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 46 | GC | 3 | Positive | 20 | 20, 20 | 0.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 47 | GC | 3 | Positive | 20 | 20, 60 | 23.094 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 48 | ESCC | 3 | Positive | 30 | 20, 50 | 15.275 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 49 | GC | 3 | Positive | 30 | 20, 30 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 50 | EAC | 3 | Positive | 50 | 20, 80 | 30.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 51 | ESCC | 3 | Positive | 60 | 20, 60 | 23.094 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 52 | GEJ | 3 | Positive | 30 | 30, 50 | 11.547 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 53 | GC | 3 | Positive | 30 | 30, 40 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 54 | GC | 3 | Positive | 30 | 30, 50 | 11.547 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 55 | EAC | 3 | Positive | 60 | 60, 60 | 0.000 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 56 | ESCC | 3 | Positive | 70 | 60, 70 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 57 | GC | 3 | Positive | 80 | 60, 80 | 11.547 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 58 | GC | 3 | Positive | 90 | 90, 100 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 59 | EAC | 3 | Positive | 100 | 90, 100 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | | 60 | ESCC | 3 | Positive | 100 | 90, 100 | 5.774 | 100% (3/3) | 100% (3/3) | 43.9-100% | GC = Gastric adenocarcinoma; GEJ = GEJ adenocarcinoma; EAC = Esophageal Adenocarcinoma; ESCC = Esophageal Squamous Cell Carcinoma; SD = Standard Deviation; CI = Confidence Interval; CPS = Combined Positive Score; n number of results; N = total number of results *Consensus result was determined based on the majority call across all test conditions **Percent positive represents the proportion of results scoring positive (CPS ≥ 1) out of the total number of results ***95% confidence intervals were calculated using the Wilson score method PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 12 of 41 {12} #### 4. External Reproducibility The reproducibility of PD-L1 IHC 28-8 pharmDx was evaluated at three external testing sites. Negative percent agreement (NPA), positive percent agreement (PPA), and overall percent agreement (OA) were computed with two-sided 95% confidence intervals using the bootstrap method. Wilson score limits were used to calculate confidence intervals for agreement parameters with point estimates equal to 100%. Reproducibility at the CPS $\geq 1$ cutoff was assessed in a two-part study: Part A assessed inter- and intra-site endpoints and Part B assessed inter- and intra-observer endpoints. In Part A, inter-site reproducibility was assessed with 66 gastric, GEJ, and esophageal carcinoma specimens. Specimens were stained at three sites over five non-consecutive days using one reagent lot. One observer at each site evaluated the set of specimens for each day. The specimens were pre-qualified at Agilent to represent the full PD-L1 expression range, and more than 20% of specimens were in the near-cutoff range around CPS $\geq 1$. Unstained specimen sections were blinded and randomized prior to processing at each of the three external sites. Stained slides were then assessed for inter-site and intra-site reproducibility. Part B assessed inter-observer and intra-observer reproducibility using 74 gastric, GEJ, and esophageal carcinoma specimens that spanned the full expression range of PD-L1. These specimens were stained, then blinded and randomized at the Agilent facility, and shipped to the three external reproducibility sites for pathologist assessment. At each of the external sites, all IHC tests were interpreted in a blinded and randomized fashion by certified clinical pathologists. Pathologists determined positive or negative results based on the CPS $\geq 1$ cutoff, with a minimum washout period of 14 days between evaluations. The results of the reproducibility study are summarized in Table 5. Per-specimen reproducibility results are presented in Tables 6 and 7. **Table 5. Summary of Reproducibility (three sites) for CPS $\geq 1$** | Reproducibility Study | Study Design | % Agreement (95% CI) | | --- | --- | --- | | Inter-Site | Each of 66 specimens of gastric, GEJ, and esophageal carcinoma (32 PD-L1-negative and 34 PD-L1-positive), spanning the full range of PD-L1 IHC expression, was tested on five non-consecutive days. Inter-site analysis was performed across three sites, yielding a total of 450 comparisons to consensus (majority call). | NPA 96.2 (92.9, 98.8) PPA 99.2 (98.2, 100.0) OA 97.8 (96.2, 99.1) | | Intra-Site | Each of 66 specimens of gastric, GEJ, and esophageal carcinoma (32 PD-L1-negative and 34 PD-L1-positive), spanning the full range of PD-L1 IHC expression, was tested on five non-consecutive days. Intra-site analysis was performed within three | NPA 98.1 (96.6, 99.4) PPA 99.0 (97.9, 99.8) OA 98.6 (97.7, 99.4) | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 13 of 41 {13} | Reproducibility Study | Study Design | % Agreement (95% CI) | | --- | --- | --- | | | sites, yielding a total of 990 comparisons to consensus (majority call). | | | Inter-Observer | Scoring of 74 specimens of gastric, GEJ, and esophageal carcinoma (36 PD-L1-negative and 38 PD-L1-positive), spanning the full range of PD-L1 IHC expression, was performed by three pathologists, one at each of three study sites, on three non-consecutive days. Inter-observer analysis was performed across three sites, yielding a total of 666 comparisons to consensus (majority call). | NPA 88.6 (83.3, 93.2) PPA 96.8 (93.0, 100.0) OA 92.8 (89.5, 95.6) | | Intra-Observer | Scoring of 74 specimens of gastric, GEJ, and esophageal carcinoma (36 PD-L1-negative and 38 PD-L1-positive), spanning the full range of PD-L1 IHC expression, was performed by three pathologists, one at each of three study sites, on three non-consecutive days. Intra-observer analysis was performed within three sites, yielding a total of 666 comparisons to consensus (majority call). | NPA 97.3 (95.3, 99.0) PPA 98.4 (97.0, 99.5) OA 97.9 (96.8, 98.8) | NPA= Negative Percent Agreement; PPA= Positive Percent Agreement; OA=Overall Percent Agreement; CPS=Combined Positive Score To accurately assess PD-L1 expression status, challenging borderline cases (slides) with scores near the CPS ≥ 1 cutoff (CPS > 0 and < 10) should be reviewed by an additional pathologist and the reported results should be based on a consensus score. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 14 of 41 {14} Table 6. Inter-Site Reproducibility Results per Specimen (Part A) | Specimen | Subtype | N | Consensus Result* | Median (CPS) | Range (CPS) | Standard Deviation** | | | Percent Positive (n/N) | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | | | | Inter-Site | Inter-Round | Total | Site 1 | Site 2 | Site 3 | All Sites | | 1 | GC | 15 | Negative | 0 | 0, <1 | 0.223 | 0.003 | 0.258 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 2 | GC | 15 | Negative | 0 | 0, <1 | 0.109 | 0.000 | 0.109 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 3 | EAC | 15 | Negative | <1 | 0, <1 | 0.091 | 0.003 | 0.183 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 4 | EAC | 15 | Negative | <1 | 0, <1 | 0.000 | 0.000 | 0.000 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 5 | GEJ | 15 | Negative | <1 | 0, <1 | 0.000 | 0.000 | 0.000 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 6 | GC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 7 | GC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 8 | GC | 15 | Negative | <1 | 0, <1 | 0.258 | 0.002 | 0.289 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 9 | GC | 15 | Negative | <1 | 0, <1 | 0.091 | 0.003 | 0.183 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 10 | GC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 11 | ESCC | 15 | Negative | <1 | 0, <1 | 0.041 | 0.000 | 0.208 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 12 | ESCC | 15 | Negative | <1 | 0, <1 | 0.258 | 0.002 | 0.289 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 13 | ESCC | 15 | Negative | <1 | 0, <1 | 0.158 | 0.004 | 0.224 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 14 | ESCC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 15 | GEJ | 15 | Negative | <1 | 0, <1 | 0.194 | 0.065 | 0.266 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 16 | GC | 15 | Negative | <1 | 0, <1 | 0.158 | 0.004 | 0.224 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 17 | ESCC | 15 | Negative | <1 | 0, <1 | 0.158 | 0.004 | 0.224 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 18 | ESCC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 19 | ESCC | 15 | Negative | <1 | 0, <1 | 0.091 | 0.003 | 0.183 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 20 | ESCC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 21 | ESCC | 15 | Negative | <1 | 0, <1 | 0.258 | 0.002 | 0.289 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 22 | GC | 15 | Negative | <1 | 0, <1 | 0.158 | 0.004 | 0.224 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 23 | GC | 15 | Negative | <1 | 0, <1 | 0.241 | 0.006 | 0.288 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 24 | ESCC | 15 | Negative | <1 | 0, <1 | 0.002 | 0.001 | 0.129 | 0% (0/5) | 0% (0/5) | 0% (0/5) | 0% (0/15) | | 25 | EAC | 15 | Negative | <1 | 0, 1 | 0.000 | 0.053 | 0.230 | 0% (0/5) | 20% (1/5) | 0% (0/5) | 6.7% (1/15) | | 26 | GEJ | 15 | Negative | <1 | 0, 1 | 0.065 | 0.144 | 0.266 | 0% (0/5) | 20% (1/5) | 0% (0/5) | 6.7% (1/15) | | 27 | GC | 15 | Negative | <1 | 0, 1 | 0.065 | 0.065 | 0.194 | 0% (0/5) | 20% (1/5) | 0% (0/5) | 6.7% (1/15) | | 28 | GC | 15 | Negative | <1 | 0, 1 | 0.065 | 0.065 | 0.233 | 0% (0/5) | 0% (0/5) | 20% (1/5) | 6.7% (1/15) | | 29 | GC | 15 | Negative | <1 | 0, 1 | 0.129 | 0.091 | 0.242 | 0% (0/5) | 40% (2/5) | 0% (0/5) | 13.3% (2/15) | | 30 | GC | 15 | Negative | <1 | <1, 2 | 0.000 | 0.000 | 0.399 | 0% (0/5) | 20% (1/5) | 20% (1/5) | 13.3% (2/15) | | 31 | EAC | 15 | Negative | <1 | <1, 1 | 0.194 | 0.065 | 0.266 | 0% (0/5) | 80% (4/5) | 20% (1/5) | 33.3% (5/15) | | 32 | GEJ | 15 | Negative | <1 | <1, 1 | 0.109 | 0.000 | 0.109 | 0% (0/5) | 100% (5/5) | 0% (0/5) | 33.3% (5/15) | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 15 of 41 {15} | 33 | GC | 15 | Positive | 3 | <1, 10 | 2.790 | 0.001 | 3.426 | 60% (3/5) | 100% (5/5) | 100% (5/5) | 86.7% (13/15) | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 34 | GEJ | 15 | Positive | 3 | <1, 5 | 1.639 | 0.000 | 1.863 | 80% (4/5) | 100% (5/5) | 100% (5/5) | 93.3% (14/15) | | 35 | EAC | 15 | Positive | 10 | <1, 40 | 6.683 | 4.703 | 10.755 | 100% (5/5) | 100% (5/5) | 80% (4/5) | 93.3% (14/15) | | 36 | EAC | 15 | Positive | 3 | 1, 10 | 2.099 | 0.000 | 2.764 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 37 | ESCC | 15 | Positive | 3 | 1, 5 | 1.449 | 0.000 | 1.807 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 38 | ESCC | 15 | Positive | 4 | 1, 15 | 1.073 | 1.910 | 4.566 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 39 | GEJ | 15 | Positive | 5 | 1, 17 | 0.014 | 2.091 | 4.800 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 40 | ESCC | 15 | Positive | 5 | 1, 17 | 2.149 | 3.686 | 5.136 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 41 | GEJ | 15 | Positive | 6 | 1, 80 | 17.867 | 7.871 | 25.368 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 42 | ESCC | 15 | Positive | 8 | 2, 70 | 12.686 | 0.000 | 18.932 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 43 | GC | 15 | Positive | 13 | 2.5, 50 | 5.312 | 0.000 | 12.198 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 44 | EAC | 15 | Positive | 30 | 3, 50 | 9.110 | 2.693 | 12.107 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 45 | GC | 15 | Positive | 13 | 5, 25 | 2.376 | 0.000 | 6.456 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 46 | GEJ | 15 | Positive | 20 | 5, 60 | 13.298 | 1.106 | 18.792 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 47 | GC | 15 | Positive | 11 | 7, 40 | 0.000 | 5.792 | 9.227 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 48 | GC | 15 | Positive | 20 | 8, 40 | 6.664 | 0.005 | 9.255 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 49 | EAC | 15 | Positive | 12 | 9, 50 | 11.714 | 0.000 | 14.924 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 50 | GC | 15 | Positive | 15 | 9, 70 | 24.922 | 0.000 | 25.595 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 51 | ESCC | 15 | Positive | 15 | 10, 70 | 20.641 | 3.652 | 24.308 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 52 | GC | 15 | Positive | 20 | 10, 40 | 4.372 | 0.000 | 9.139 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 53 | EAC | 15 | Positive | 21 | 10, 80 | 18.625 | 3.014 | 22.715 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 54 | GEJ | 15 | Positive | 25 | 10, 60 | 0.000 | 0.000 | 12.993 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 55 | EAC | 15 | Positive | 25 | 10, 65 | 14.335 | 2.893 | 19.864 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 56 | EAC | 15 | Positive | 35 | 10, 70 | 20.736 | 0.000 | 22.683 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 57 | ESCC | 15 | Positive | 40 | 10, 75 | 23.702 | 0.000 | 27.557 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 58 | GC | 15 | Positive | 40 | 13, 100 | 36.700 | 0.000 | 37.374 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 59 | EAC | 15 | Positive | 30 | 17, 60 | 8.491 | 0.001 | 13.262 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 60 | GC | 15 | Positive | 60 | 23, 100 | 25.230 | 0.008 | 26.783 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 61 | GC | 15 | Positive | 65 | 30, 90 | 14.374 | 0.000 | 19.761 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 62 | GC | 15 | Positive | 80 | 40, 100 | 15.054 | 0.000 | 21.567 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 63 | ESCC | 15 | Positive | 80 | 40, 95 | 14.742 | 5.686 | 19.990 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 64 | GC | 15 | Positive | 85 | 60, 100 | 12.113 | 0.000 | 14.617 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 65 | EAC | 15 | Positive | 100 | 80, 100 | 1.709 | 3.096 | 6.677 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | | 66 | GEJ | 15 | Positive | 100 | 80, 100 | 1.581 | 4.183 | 6.519 | 100% (5/5) | 100% (5/5) | 100% (5/5) | 100% (15/15) | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 16 of 41 {16} GC = Gastric adenocarcinoma; GEJ = GEJ adenocarcinoma; EAC = Esophageal Adenocarcinoma; ESCC = Esophageal Squamous Cell Carcinoma; SD = Standard Deviation; CI = Confidence Interval; CPS = Combined Positive Score; n = number of results; N = total number of results *Consensus result was determined based on the majority call across all test conditions **Percent positive represents the proportion of results scoring positive (CPS ≥ 1) out of the total number of results ***95% confidence intervals were calculated using the Wilson score method Table 7. Inter-Observer Reproducibility Results per Specimen (Part B) | Specimen | Subtype | N | Consensus Result* | Median (CPS) | Range (CPS) | Standard Deviation | | | Percent Positive (n/N) | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | | | | Inter-Observer | Inter-Read | Total | Observer 1 | Observer 2 | Observer 3 | All Observers | | 1 | GEJ | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 2 | GEJ | 9 | Negative | <1 | 0, <1 | 0.144 | 0.000 | 0.144 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 3 | EAC | 9 | Negative | <1 | 0, <1 | 0.167 | 0.167 | 0.236 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 4 | GEJ | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 5 | ESCC | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 6 | EAC | 9 | Negative | <1 | 0, <1 | 0.144 | 0.000 | 0.144 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 7 | GEJ | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 8 | EAC | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 9 | ESCC | 9 | Negative | <1 | 0, <1 | 0.144 | 0.000 | 0.144 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 10 | ESCC | 9 | Negative | <1 | 0, <1 | 0.144 | 0.000 | 0.144 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 11 | GC | 9 | Negative | <1 | 0, <1 | 0.167 | 0.167 | 0.236 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 12 | ESCC | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 13 | ESCC | 9 | Negative | <1 | 0, <1 | 0.236 | 0.167 | 0.289 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 14 | ESCC | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 15 | ESCC | 9 | Negative | <1 | 0, <1 | 0.000 | 0.167 | 0.167 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 16 | GC | 9 | Negative | <1 | 0, <1 | 0.144 | 0.000 | 0.144 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 17 | ESCC | 9 | Negative | <1 | 0, 2 | 0.000 | 0.546 | 0.546 | 0% (0/3) | 33.3% (1/3) | 0% (0/3) | 11.1% (1/9) | | 18 | GC | 9 | Negative | <1 | 0, 2 | 0.000 | 0.583 | 0.583 | 0% (0/3) | 33.3% (1/3) | 0% (0/3) | 11.1% (1/9) | | 19 | GC | 9 | Negative | <1 | 0, 1 | 0.000 | 0.250 | 0.250 | 0% (0/3) | 33.3% (1/3) | 0% (0/3) | 11.1% (1/9) | | 20 | EAC | 9 | Negative | <1 | 0, 4 | 0.000 | 1.193 | 1.193 | 0% (0/3) | 66.7% (2/3) | 0% (0/3) | 22.2% (2/9) | | 21 | GEJ | 9 | Negative | <1 | <1, <1 | 0.000 | 0.000 | 0.000 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 22 | GC | 9 | Negative | <1 | <1, <1 | 0.000 | 0.000 | 0.000 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 23 | GC | 9 | Negative | <1 | <1, <1 | 0.000 | 0.000 | 0.000 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 24 | GC | 9 | Negative | <1 | <1, <1 | 0.000 | 0.000 | 0.000 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 25 | GC | 9 | Negative | <1 | <1, <1 | 0.000 | 0.000 | 0.000 | 0% (0/3) | 0% (0/3) | 0% (0/3) | 0% (0/9) | | 26 | GEJ | 9 | Negative | <1 | <1, 1 | 0.167 | 0.167 | 0.236 | 0% (0/3) | 66.7% (2/3) | 0% (0/3) | 22.2% (2/9) | | 27 | ESCC | 9 | Negative | <1 | <1, 1 | 0.167 | 0.167 | 0.236 | 0% (0/3) | 66.7% (2/3) | 0% (0/3) | 22.2% (2/9) | | 28 | GC | 9 | Negative | <1 | <1, 1 | 0.000 | 0.220 | 0.220 | 0% (0/3) | 33.3% (1/3) | 33.3% (1/3) | 22.2% (2/9) | | 29 | ESCC | 9 | Negative | <1 | <1, 1 | 0.167 | 0.167 | 0.236 | 0% (0/3) | 66.7% (2/3) | 0% (0/3) | 22.2% (2/9) | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 17 of 41 {17} | 30 | EAC | 9 | Negative | <1 | <1, 3 | 0.799 | 0.577 | 0.986 | 0% (0/3) | 100% (3/3) | 0% (0/3) | 33.3% (3/9) | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 31 | GEJ | 9 | Negative | <1 | <1, 2 | 0.441 | 0.333 | 0.553 | 0% (0/3) | 100% (3/3) | 0% (0/3) | 33.3% (3/9) | | 32 | GC | 9 | Negative | <1 | <1, 5 | 1.555 | 0.882 | 1.787 | 0% (0/3) | 100% (3/3) | 0% (0/3) | 33.3% (3/9) | | 33 | GC | 9 | Negative | <1 | <1, 2 | 0.441 | 0.333 | 0.553 | 0% (0/3) | 100% (3/3) | 0% (0/3) | 33.3% (3/9) | | 34 | EAC | 9 | Negative | <1 | <1, 3 | 0.491 | 0.687 | 0.844 | 0% (0/3) | 100% (3/3) | 33.3% (1/3) | 44.4% (4/9) | | 35 | EAC | 9 | Negative | <1 | <1, 2 | 0.236 | 0.601 | 0.645 | 0% (0/3) | 100% (3/3) | 33.3% (1/3) | 44.4% (4/9) | | 36 | GEJ | 9 | Negative | <1 | <1, 3 | 0.745 | 0.601 | 0.957 | 0% (0/3) | 100% (3/3) | 33.3% (1/3) | 44.4% (4/9) | | 37 | EAC | 9 | Positive | 1 | <1, 3 | 1.147 | 0.373 | 1.206 | 0% (0/3) | 100% (3/3) | 66.7% (2/3) | 55.6% (5/9) | | 38 | ESCC | 9 | Positive | 1 | <1, 4 | 0.561 | 1.067 | 1.206 | 0% (0/3) | 100% (3/3) | 66.7% (2/3) | 55.6% (5/9) | | 39 | EAC | 9 | Positive | 3 | <1, 6 | 2.217 | 0.667 | 2.315 | 0% (0/3) | 100% (3/3) | 100% (3/3) | 66.7% (6/9) | | 40 | GC | 9 | Positive | 11 | 5, 15 | 3.448 | 3.127 | 4.655 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 41 | GEJ | 9 | Positive | 13 | 6, 30 | 4.970 | 6.342 | 8.058 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 42 | GC | 9 | Positive | 15 | 8, 50 | 0.000 | 13.011 | 13.011 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 43 | ESCC | 9 | Positive | 16 | 9, 25 | 6.546 | 2.749 | 7.100 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 44 | EAC | 9 | Positive | 20 | 10, 25 | 5.260 | 3.528 | 6.333 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 45 | EAC | 9 | Positive | 20 | 10, 40 | 0.000 | 9.997 | 9.997 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 46 | ESCC | 9 | Positive | 25 | 10, 31 | 0.000 | 7.550 | 7.550 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 47 | GC | 9 | Positive | 25 | 10, 50 | 17.816 | 6.446 | 18.946 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 48 | GEJ | 9 | Positive | 30 | 10, 65 | 20.262 | 6.254 | 21.205 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 49 | GC | 9 | Positive | 35 | 10, 40 | 8.472 | 8.699 | 12.143 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 50 | EAC | 9 | Positive | 25 | 12, 35 | 3.203 | 7.454 | 8.113 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 51 | GC | 9 | Positive | 25 | 12, 30 | 4.823 | 4.955 | 6.915 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 52 | EAC | 9 | Positive | 27 | 12, 30 | 3.512 | 5.333 | 6.386 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 53 | GEJ | 9 | Positive | 25 | 14, 40 | 3.585 | 7.542 | 8.351 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 54 | GEJ | 9 | Positive | 75 | 15, 80 | 5.225 | 23.627 | 24.197 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 55 | GC | 9 | Positive | 26 | 19, 50 | 13.784 | 4.738 | 14.575 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 56 | EAC | 9 | Positive | 41 | 20, 80 | 22.476 | 13.671 | 26.307 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 57 | GC | 9 | Positive | 40 | 21, 100 | 0.000 | 27.597 | 27.597 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 58 | GEJ | 9 | Positive | 40 | 22, 65 | 13.910 | 10.044 | 17.157 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 59 | EAC | 9 | Positive | 29 | 23, 50 | 13.387 | 3.887 | 13.940 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 60 | GEJ | 9 | Positive | 33 | 23, 40 | 6.272 | 3.480 | 7.172 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 61 | ESCC | 9 | Positive | 30 | 28, 38 | 0.000 | 2.819 | 2.819 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 62 | GEJ | 9 | Positive | 40 | 28, 50 | 7.071 | 4.333 | 8.293 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 63 | GEJ | 9 | Positive | 53 | 33, 70 | 13.241 | 10.493 | 16.895 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 64 | ESCC | 9 | Positive | 75 | 46, 85 | 14.376 | 7.965 | 16.435 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 18 of 41 {18} | 65 | ESCC | 9 | Positive | 85 | 51, 90 | 7.520 | 12.901 | 14.933 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | 66 | GEJ | 9 | Positive | 100 | 67, 100 | 11.499 | 6.658 | 13.287 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 67 | EAC | 9 | Positive | 98 | 70, 100 | 8.119 | 6.823 | 10.606 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 68 | ESCC | 9 | Positive | 95 | 72, 100 | 9.841 | 5.467 | 11.258 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 69 | GC | 9 | Positive | 95 | 80, 100 | 7.175 | 3.859 | 8.147 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 70 | GEJ | 9 | Positive | 100 | 90, 100 | 0.001 | 3.333 | 3.333 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 71 | GC | 9 | Positive | 100 | 90, 100 | 0.001 | 3.333 | 3.333 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 72 | ESCC | 9 | Positive | 100 | 90, 100 | 0.001 | 3.333 | 3.333 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 73 | ESCC | 9 | Positive | 100 | 90, 100 | 0.001 | 3.333 | 3.333 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | | 74 | EAC | 9 | Positive | 100 | 98, 100 | 0.000 | 0.667 | 0.667 | 100% (3/3) | 100% (3/3) | 100% (3/3) | 100% (9/9) | GC = Gastric adenocarcinoma; GEJ = GEJ adenocarcinoma; EAC = Esophageal Adenocarcinoma; ESCC = Esophageal Squamous Cell Carcinoma; SD = Standard Deviation; CI = Confidence Interval; CPS = Combined Positive Score; n = number of results; N = total number of results *Consensus result was determined based on the majority call across all test conditions **Percent positive represents the proportion of results scoring positive (CPS ≥ 1) out of the total number of results ***95% confidence intervals were calculated using the Wilson score method ### 5. Robustness Studies The robustness of PD-L1 IHC 28-8 pharmDx staining was evaluated by testing the assay on gastric, GEJ, and esophageal carcinoma specimens under various laboratory conditions. Conditions tested included: - Tissue section thickness (3, 4, and 5 μm) - Target Retrieval Solution temperature (97 °C, 95 °C, and 99 °C) - Target Retrieval time (20, 18, and 22 minutes) - Target Retrieval Solution pH (pH 6.1, 5.9, and 6.3) - Target Retrieval Solution re-use (up to 3 re-uses) Approximately 150 gastric, GEJ, and esophageal carcinoma specimens spanning the range of PD-L1 expression, including specimens near the CPS ≥ 1 cutoff, were included for each condition tested. All specimens were stained, then blinded and randomized prior to evaluation of PD-L1 expression status. Staining performance was evaluated using the CPS scoring method. No significant difference in results was observed for any of the experimental conditions tested. Pre-specified acceptance criteria of 85% at the lower bound of a two-sided 95% confidence interval for PPA, NPA, and OPA were met in these studies. ### 6. Impact of Tumor Heterogeneity The objective of the studies described below was to investigate whether tumor heterogeneity affects PD-L1 IHC staining results with PD-L1 IHC 28-8 pharmDx. #### a. Primary vs. Metastatic Tumor Tissues Matched primary versus metastatic blocks were obtained from 27 subjects (gastric, GEJ, and esophageal carcinoma) and evaluated by PD-L1 IHC 28-8 pharmDx. In 21 PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 19 of 41 {19} of 27 matched pairs, the diagnostic outcome was identical for primary and metastatic specimens at CPS ≥ 1. Six matched pairs showed discordant results at CPS ≥ 1. # *b. Multiple FFPE Blocks from the Same Cases* Multiple blocks (2 or more) from each of 38 specimens (gastric, GEJ, and esophageal carcinoma) obtained from the same tumor were evaluated. In 36 of 38 sets of intra-subject specimens, the diagnostic outcomes were identical for all specimens at CPS ≥ 1. Two cases showed discordant results at CPS ≥ 1. ## 7. Stability Testing No changes have been made to the device design, including reagent formulation or kit configuration, since approval of P150025. The real-time stability study results support the stability of PD-L1 IHC 28-8 pharmDx reagents as follows: # *a. PD-L1 IHC 28-8 pharmDx Stability* # Finished Good Shelf Life 12 months at 2-8 °C # In-Use/On-Board Stability Testing Fifteen cycles to room temperature # Working/Reconstituted Stability Testing DAB Substrate-Chromogen Solution: 5 Days at 2-8 °C, protected from Light # Target Retrieval Solution Five days at room temperature in a PT Link with up to 3 uses for 3-in-1 Pretreatment # *b. Gastric, GEJ, and Esophageal Carcinoma FFPE Cut Section Stability* Real-time cut section stability studies were conducted to evaluate the shelf life of cut tissue sections of gastric, GEJ, and esophageal carcinoma FFPE blocks using PD-L1 IHC 28-8 pharmDx when stored in the dark at 2-8 °C or 25 °C. Based on the results of the study, tissue sections of each specimen type are stable for up to 4 months when stored in the dark at 2–8 °C or at room temperature (up to 25 °C). ## B. Animal Studies None ## C. Additional Studies None ---PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 20 of 41 {20} # X. SUMMARY OF PRIMARY CLINICAL STUDIES # A. Clinical Performance Evaluation: Esophageal Squamous Cell Carcinoma (ESCC) CHECKMATE-648 (CA209648) The clinical performance of PD-L1 IHC 28-8 pharmDx was evaluated in CHECKMATE-648 to establish a reasonable assurance of safety and effectiveness of the device for identifying patients diagnosed with unresectable advanced, recurrent, or metastatic previously untreated ESCC for treatment with OPDIVO (nivolumab) plus YERVOY (ipilimumab) or OPDIVO combined with fluorouracil and cisplatin. Data from this clinical study were the basis for the sPMA approval decision. The clinical study, conducted with intravenous OPDIVO (nivolumab), also served as the primary evidence supporting the effectiveness of OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy) for subcutaneous use of nivolumab in combination with fluoropyrimidine and platinum-containing chemotherapy for the first-line treatment of unresectable advanced or metastatic ESCC. The indication for OPDIVO QVANTIG for this patient population is further supported by evidence from the CHECKMATE-67T trial, which demonstrated comparable pharmacokinetics and safety profiles between the subcutaneous and intravenous formulations of nivolumab (see the OPDIVO QVANTIG product label for safety, clinical pharmacology, and effectiveness). # 1. Study Design CHECKMATE-648 was a randomized, open-label, multicenter Phase 3 study comparing OPDIVO (nivolumab) plus YERVOY (ipilimumab) or OPDIVO combined with fluorouracil and cisplatin versus standard of care (fluorouracil plus cisplatin) in previously untreated unresectable advanced, recurrent, or metastatic ESCC. Patients were randomized in a 1:1:1 ratio to one of the following treatments (Arms A, B, and C): - Arm A (OPDIVO + ipilimumab): nivolumab 3 mg/kg every 2 weeks (Q2W) intravenously (IV) + ipilimumab 1 mg/kg every 6 weeks (Q6W) IV - Arm B (OPDIVO + chemotherapy): nivolumab 240 mg Q2W IV + fluorouracil 800 mg/m²/day IV on Day 1 through Day 5 + cisplatin 80 mg/m² IV on Day 1 of a 4-week cycle - Arm C (chemotherapy): fluorouracil 800 mg/m²/day IV on Day 1 through Day 5 + cisplatin 80 mg/m² IV on Day 1 of a 4-week cycle At randomization, patients were stratified according to tumor cell PD-L1 status: ≥ 1% vs. < 1% (including indeterminate). The proportions of subjects with or without tumor cell PD-L1 expression were monitored as needed to ensure that the sample size of randomized subjects with tumor cell PD-L1 expression ≥ 1% was adequate for analysis (i.e., approximately 50% of all randomized subjects). Clinical validation of the device in this study was limited to the population of patients with PD-L1 CPS ≥ 1 treated with OPDIVO (nivolumab) combined with YERVOY (ipilimumab) or OPDIVO combined with fluorouracil plus cisplatin. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 21 of 41 {21} All randomized subjects had their PD-L1 stained slides (or images for the samples originally stained in China) re-scored for Combined Positive Score (CPS). # a. Clinical Inclusion and Exclusion Criteria The key inclusion and exclusion criteria were as follows. # Key Inclusion Criteria: - Histologically confirmed squamous cell carcinoma or adenosquamous cell carcinoma (predominant squamous differentiation) of esophagus (per AJCC 7th edition). - Have unresectable advanced, recurrent, or metastatic ESCC (per AJCC 7th edition). - Not amenable to curative approaches such as definitive chemoradiation and/or surgery. - No prior systemic anticancer therapy given as primary therapy for advanced or metastatic disease. Prior adjuvant, neoadjuvant, or definitive chemotherapy, radiotherapy, or chemoradiotherapy for ESCC was permitted if given as part of a curative-intent regimen and completed before enrollment. - ECOG Performance Status of 0 or 1. - At least one measurable lesion by CT or MRI per RECIST 1.1 criteria; radiographic tumor assessment performed within 28 days prior to randomization. - Provided either a newly obtained or archival tissue sample for biomarker analyses. - Have an evaluable PD-L1 expression classification of ≥ 1% or <1% (or indeterminate) as determined by the central lab. # Key Exclusion Criteria: - Have not recovered from the effects of major surgery or significant traumatic injury at least 14 days prior to randomization. - Have a prior malignancy requiring active treatment within the previous 3 years except for locally curable cancers that had been apparently cured, such as basal or squamous cell skin cancer, superficial bladder cancer, or carcinoma in situ of the prostate, cervix, or breast. - Have active, known, or suspected autoimmune disease. Subjects with Type I diabetes mellitus, residual hypothyroidism due to autoimmune thyroiditis only requiring hormone replacement, skin disorders (such as vitiligo, psoriasis, or alopecia) not requiring systemic treatment were permitted to enroll. - Have a condition requiring systemic treatment with either corticosteroids (> 10 mg daily prednisone equivalent) or other immunosuppressive medications within 14 days of start of study treatment. - Have received prior treatment with an anti-PD-1, anti-PD-L1, anti-PD-L2, anti-CD137, or anti-CTLA-4 antibody, or any other antibody or drug specifically targeting T-cell co-stimulation or checkpoint pathways. # b. Follow-up Schedule Treatment with OPDIVO (nivolumab) in Arms A and B was given for up to 24 months in the absence of disease progression or unacceptable toxicity. Chemotherapy (Arms B and C) was given as per the study dosing schedule until disease progression, unacceptable toxicity, or other reasons specified in the protocol. The study had two safety follow-up (FU) visits: PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 22 of 41 {22} 1) FU1 = 30 days from last dose ($\pm$ 7 days) or coinciding with the date of discontinuation ($\pm$ 7 days) if the date of discontinuation was greater than 35 days after last dose, and 2) FU2 = 84 days from FU1 ($\pm$ 7 days). After completion of the first two follow-up visits, subjects were followed every 3 months ($\pm$ 14 days) for overall survival. Tumor assessments per RECIST v1.1 were conducted every 6 weeks up to and including week 48, then every 12 weeks thereafter until disease progression (unless treatment beyond progressive disease, or PD, was permitted), or the subject discontinued the study, whichever came first. Subjects who discontinued study treatment for reasons other than PD continued to have tumor assessments until their disease progressed or they withdrew consent. # c. Clinical Endpoints Clinical study CA209648 (CHECKMATE-648) was designed to evaluate the efficacy and safety of OPDIVO-based regimens in previously untreated ESCC, with the following objectives. For the primary efficacy analysis, overall survival (OS) and progression-free survival (PFS) as assessed by BICR were to be compared between OPDIVO + ipilimumab (Arm A) and chemotherapy alone (Arm C), and between OPDIVO + chemotherapy (Arm B) and chemotherapy alone (Arm C) in all subjects with tumor cell PD-L1 expression $\geq 1\%$. For the key secondary and exploratory efficacy analyses, PFS per BICR and OS were to be compared in all randomized subjects between OPDIVO + ipilimumab (Arm A) and chemotherapy alone (Arm C), and between OPDIVO + chemotherapy (Arm B) and chemotherapy alone (Arm C). ORR as assessed by BICR was determined for each treatment group, both in subjects with PD-L1-expressing tumors and in all randomized subjects. Duration of response (DOR) as assessed by BICR was evaluated in subjects who achieved partial response (PR) or complete response (CR), in subjects with PD-L1-expressing tumors, and in each treatment group of all randomized subjects. # d. Assay Cut-Off Selection On September 26, 2024, FDA convened an Oncologic Drugs Advisory Committee (ODAC) to discuss the risk-benefit assessment of the use of immune checkpoint inhibitors (ICIs) in combination with chemotherapy for the first-line treatment of patients with unresectable or metastatic esophageal squamous cell carcinoma (ESCC) at different levels of PD-L1 protein expression. FDA asked members of the ODAC if the risk-benefit assessment was favorable for the use of these agents in patients with PD-L1 CPS $< 1$. After extensive discussion and the ODAC's agreement with FDA's position, a decision was made to limit use of these products to patients with PD-L1 CPS $\geq 1$. Subsequently, the esophageal squamous cell carcinoma indication labeling for OPDIVO (nivolumab) and OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy) was revised to PD-L1 CPS $\geq 1$. For this supplementary PMA, the data represent an exploratory analysis of subgroups at the PD-L1 CPS $\geq 1$ cutoff, per FDA's therapy label updates following ODAC recommendations. ## 2. Accountability of PMA Cohort A total of 970 patients were randomized in CheckMate-648. Among all randomized patients, 965 had quantifiable tumor cell PD-L1 expression at baseline: 322 of 325 (99.1%), 321 of 321 ---PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 23 of 41 {23} (100%), and 322 of 324 (99.4%) in the OPDIVO (nivolumab) + ipilimumab, OPDIVO + chemotherapy, and chemotherapy alone arms, respectively. PD-L1 CPS status was determined for all randomized patients by re-scoring PD-L1-stained slides or whole slide images for samples originally stained in China. Of all randomized patients, 906 (93.4%) had quantifiable PD-L1 by CPS: 297 (91.4%) in OPDIVO + ipilimumab, 305 (95.0%) in OPDIVO + chemotherapy, and 304 (93.8%) in the chemotherapy arm. Of these 906 patients, 824 (90.9%) had PD-L1 expression by CPS ≥ 1: 266 (89.6%) in OPDIVO + ipilimumab, 278 (91.1%) in OPDIVO + chemotherapy, and 280 (92.1%) in the chemotherapy arm. Accountability of the PMA cohort is provided in Table 8. Table 8. Accountability of PMA Cohort for All Randomized Patients with PD-L1 CPS ≥1 | | OPDIVO + Ipilimumab (N=266) | OPDIVO + Chemotherapy (N=278) | Chemotherapy (N=280) | Total (N=824) | | --- | --- | --- | --- | --- | | SUBJECTS WITH TUMOR SAMPLE COLLECTED AT BASELINE | 266 (100.0) | 278 (100.0) | 280 (100.0) | 824 (100.0) | | ANATOMIC LOCATION BY SITE OF COLLECTION ON SPECIMEN (1) | | | | | | PRIMARY | 209 (78.6) | 218 (78.4) | 229 (81.8) | 656 (79.6) | | METASTASIS | 57 (21.4) | 60 (21.6) | 51 (18.2) | 168 (20.4) | | BONE WITH SOFT TISSUE COMPONENT | 0 | 2 (0.7) | 2 (0.7) | 4 (0.5) | | BRAIN | 0 | 0 | 1 (0.4) | 1 (0.1) | | CHEST WALL | 1 (0.4) | 0 | 0 | 1 (0.1) | | COLON | 0 | 1 (0.4) | 2 (0.7) | 3 (0.4) | | ESOPHAGUS | 2 (0.8) | 5 (1.8) | 2 (0.7) | 9 (1.1) | | KIDNEY | 0 | 1 (0.4) | 0 | 1 (0.1) | | LIVER | 10 (3.8) | 4 (1.4) | 6 (2.1) | 20 (2.4) | | LUNG | 13 (4.9) | 15 (5.4) | 10 (3.6) | 38 (4.6) | | LYMPH NODE | 22 (8.3) | 25 (9.0) | 21 (7.5) | 68 (8.3) | | SOFT TISSUE | 1 (0.4) | 3 (1.1) | 2 (0.7) | 6 (0.7) | | STOMACH | 1 (0.4) | 2 (0.7) | 3 (1.1) | 6 (0.7) | | NOT REPORTED | 7 (2.6) | 2 (0.7) | 2 (0.7) | 11 (1.3) | | SPECIMEN TYPE (1) | | | | | | SLIDES | 190 (71.4) | 190 (68.3) | 196 (70.0) | 576 (69.9) | | TISSUE BLOCK | 76 (28.6) | 88 (31.7) | 84 (30.0) | 248 (30.1) | | BIOPSY TYPE (1) | | | | | | CORE NEEDLE BIOPSY | 96 (36.1) | 110 (39.6) | 91 (32.5) | 297 (36.0) | | EXCISIONAL BIOPSY | 59 (22.2) | 73 (26.3) | 68 (24.3) | 200 (24.3) | | INCISIONAL BIOPSY | 50 (18.8) | 43 (15.5) | 58 (20.7) | 151 (18.3) | | SURGICAL RESECTION | 26 (9.8) | 29 (10.4) | 28 (10.0) | 83 (10.1) | | OTHER | 35 (13.2) | 23 (8.3) | 35 (12.5) | 93 (11.3) | | SPECIMEN STATUS (1) | | | | | | ARCHIVAL | 200 (75.2) | 196 (70.5) | 208 (74.3) | 604 (73.3) | | FRESH | 35 (13.2) | 47 (16.9) | 37 (13.2) | 119 (14.4) | | NOT REPORT | 31 (11.7) | 35 (12.6) | 35 (12.5) | 101 (12.3) | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 24 of 41 {24} Per protocol, the tumor tissue collection was to occur prior to randomization. The specimen characteristics are from the sample handling information of the baseline specimens collected at the patient enrollment, then scored for CPS. (1) Percentages are based on the subjects with tumor tissue sample collected at baseline. ### 3. Study Population Demographics and Baseline Parameters Demographics and baseline disease characteristics were generally balanced in the CPS ≥ 1 population among the OPDIVO (nivolumab) + ipilimumab, OPDIVO + chemotherapy, and chemotherapy arms. Disease characteristics of randomized subjects with PD-L1 CPS ≥ 1 were representative of patients with unresectable, advanced, recurrent, or metastatic previously untreated ESCC. The trial population characteristics in patients with PD-L1 CPS ≥ 1 were: median age of 63 years (range: 26 to 90), 45.8% aged 65 years or older; 82.3% male, 71.2% Asian, 25.2% White, and 1.2% Black or African American. Patients had histological confirmation of squamous cell carcinoma (98.3%) or adenosquamous cell carcinoma (1.7%) in the esophagus. Baseline ECOG performance status was 0 (47%) or 1 (52.9%). Table 9. Baseline Characteristics for Study Subjects with PD-L1 CPS ≥ 1 | | OPDIVO + Ipilimumab (N=266) | OPDIVO + Chemotherapy (N=278) | Chemotherapy (N=280) | Total (N=824) | | --- | --- | --- | --- | --- | | AGE (YEARS) | | | | | | N | 266 | 278 | 280 | 824 | | Mean | 62.1 | 63.2 | 63.2 | 62.8 | | Median | 62.0 | 63.5 | 64.0 | 63.0 | | Min, Max | 28, 81 | 40, 90 | 26, 81 | 26, 90 | | SD | 9.3 | 9.2 | 8.6 | 9.0 | | < 65 | 154 (57.9) | 145 (52.2) | 148 (52.9) | 447 (54.2) | | ≥ 65 | 112 (42.1) | 133 (47.8) | 132 (47.1) | 377 (45.8) | | SEX (%) | | | | | | Male | 226 (85.0) | 217 (78.1) | 235 (83.9) | 678 (82.3) | | Female | 40 (15.0) | 61 (21.9) | 45 (16.1) | 146 (17.7) | | RACE (%) | | | | | | White | 63 (23.7) | 73 (26.3) | 72 (25.7) | 208 (25.2) | | Black or African American | 3 (1.1) | 1 (0.4) | 6 (2.1) | 10 (1.2) | | American Indian or Alaska Native | 1 (0.4) | 1 (0.4) | 0 | 2 (0.2) | | Asian Indian | 1 (0.4) | 4 (1.4) | 2 (0.7) | 7 (0.8) | | Chinese | 60 (22.6) | 66 (23.7) | 60 (21.4) | 186 (22.6) | | Japanese | 107 (40.2) | 109 (39.2) | 119 (42.5) | 335 (40.7) | | Asian Other | 23 (8.6) | 20 (7.2) | 15 (5.4) | 58 (7.0) | | Other | 8 (3.0) | 4 (1.4) | 6 (2.1) | 18 (2.2) | | GEOGRAPHIC REGION (%) | | | | | | Asia | 189 (71.1) | 197 (70.9) | 196 (70.0) | 582 (70.6) | | Non-Asia | 77 (28.9) | 81 (29.1) | 84 (30.0) | 242 (29.4) | | China | 31 (11.7) | 35 (12.6) | 35 (12.5) | 101 (12.3) | | Rest of World | 235 (88.3) | 243 (87.4) | 245 (87.5) | 723 (87.7) | | ECOG PS (PER CRF) (%) | | | | | PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 25 of 41 {25} | | OPDIVO + Ipilimumab (N=266) | OPDIVO + Chemotherapy (N=278) | Chemotherapy (N=280) | Total (N=824) | | --- | --- | --- | --- | --- | | 0 | 120 (45.1) | 134 (48.2) | 133 (47.5) | 387 (47.0) | | 1 | 146 (54.9) | 144 (51.8) | 146 (52.1) | 436 (52.9) | | Not Reported | 0 | 0 | 1 (0.4) | 1 (0.1) | | DISEASE STAGE AT INITIAL DIAGNOSIS (%) | | | | | | Stage IA | 5 (1.9) | 8 (2.9) | 1 (0.4) | 14 (1.7) | | Stage IB | 6 (2.3) | 5 (1.8) | 7 (2.5) | 18 (2.2) | | Stage IIA | 6 (2.3) | 8 (2.9) | 3 (1.1) | 17 (2.1) | | Stage IIB | 12 (4.5) | 21 (7.6) | 7 (2.5) | 40 (4.9) | | Stage IIIA | 20 (7.5) | 23 (8.3) | 21 (7.5) | 64 (7.8) | | Stage IIIB | 16 (6.0) | 12 (4.3) | 25 (8.9) | 53 (6.4) | | Stage IIIC | 27 (10.2) | 20 (7.2) | 34 (12.1) | 81 (9.8) | | Stage IV | 172 (64.7) | 180 (64.7) | 181 (64.6) | 533 (64.7) | | Not Reported | 2 (0.8) | 1 (0.4) | 1 (0.4) | 4 (0.5) | | HISTOLOGICAL CLASSIFICATION AT INITIAL DIAGNOSIS (%) | | | | | | Squamous Cell Carcinoma | 264 (99.2) | 271 (97.5) | 275 (98.2) | 810 (98.3) | | Adenosquamous Carcinoma | 2 (0.8) | 7 (2.5) | 5 (1.8) | 14 (1.7) | | Other | 0 | 0 | 0 | 0 | ### 4. Safety and Effectiveness Results #### a. Safety Results As an in vitro diagnostic test, PD-L1 IHC 28-8 pharmDx involves testing on FFPE ESCC specimens. These tissues are routinely removed as part of the practice of medicine for the diagnosis of cancer by pathologists. Removal of these tissues, therefore, presents no additional safety hazard to the patient being tested. No device-related adverse events occurred during the study. The safety of OPDIVO (nivolumab) in combination with ipilimumab or chemotherapy was evaluated in the clinical study (see the OPDIVO label). No meaningful differences in adverse events were observed based on tumor cell PD-L1 expression level in all treated subjects. Please refer to Drugs@FDA for complete safety information on OPDIVO and OPDIVO QVANTIG (nivolumab and hyaluronidase-nvhy). #### b. Effectiveness Results Analysis of overall survival (OS) in ESCC patients with PD-L1 CPS ≥ 1 tumors showed a clinically meaningful improvement in survival benefit for patients randomized to OPDIVO (nivolumab) in combination with chemotherapy or ipilimumab, compared to chemotherapy alone. Table 10 summarizes the key efficacy measures for patients with ESCC with CPS ≥ 1. Figure 1 shows the Kaplan-Meier overall survival curves for ESCC patients with CPS ≥ 1. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 26 of 41 {26} Table 10. Summary of CHECKMATE-648 Efficacy Results in Participants with PD-L1 CPS ≥1 | | OPDIVO with Cisplatin and Fluorouracil (n=278) | OPDIVO and Ipilimumab (n=266) | Cisplatin and Fluorouracil (n=280) | | --- | --- | --- | --- | | Overall Survival | | | | | Deaths (%) | 177 (64) | 179 (67) | 205 (73) | | Median (months) (95% CI) | 13.8 (12.0, 16.1) | 12.8 (10.9, 15.5) | 9.8 (8.8, 11.6) | | Hazard Ratio (95% CI)b | 0.69 (0.57, 0.85) | 0.76 (0.62, 0.93) | – | | p-valueNT | 0.0003 | 0.0086 | – | | Progression-Free Survival | | | | | Disease Progression or Death (%) | 201 (72) | 206 (77) | 184 (66) | | Median (months) (95% CI) | 5.8 (5.5, 7.0) | 2.8 (2.6, 4.2) | 5.6 (4.2, 5.9) | | Hazard Ratio (95% CI)b | 0.79 (0.65, 0.97) | 1.21 (0.98, 1.48) | – | | p-valueNT | 0.0247 | 0.0713 | – | | Overall Response Rate, n (%)a | 135 (49) | 74 (28) | 76 (27) | | (95% CI) | (42.5, 54.6) | (22.5, 33.6) | (22.0, 32.8) | | Complete Response (%) | 39 (14%) | 32 (12.0%) | 18 (6.4%) | | Partial Response (%) | 96 (34.5%) | 42 (15.8%) | 58 (20.7%) | | Duration of Response (months)a | | | | | Median (95% CI) | 8.2 (6.7, 11.1) | 11.8 (7.1, 23.6) | 6.9 (5.7, 8.2) | | Range | 1.4+, 35.9+ | 1.4+, 34.5+ | 1.4+, 31.8+ | a Assessed by BICR. b Based on stratified Cox proportional hazard model. Hazard ratios are reported for each OPDIVO containing arm compared to chemotherapy within each analysis population. NT: Not evaluated for statistical significance as per pre-specified hierarchical testing procedure. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 27 of 41 {27} ![img-0.jpeg](img-0.jpeg) Figure 1. Overall Survival of All Randomized Subjects with PD-L1 CPS ≥1 Table 11 below summarizes the key efficacy measures for patients with ESCC and CPS ≥ 1, excluding subjects from China. The exclusion of Chinese subjects did not alter the efficacy conclusions in the CPS ≥ 1 population; OS and PFS results were consistent with those observed in the overall randomized CPS ≥ 1 population. PMA P150025/S018: FDA Summary of Safety and Effectiveness Data 28 of 41 {28} Table 11. Summary of CM-648 Key Efficacy Results in All Randomized CPS ≥ 1 Subjects Excluding Subjects from China | | OPDIVO with Cisplatin and Fluorouracil (n=243) | OPDIVO and Ipilimumab (n=235) | Cisplatin and Fluorouracil (n=245) | | --- | --- | --- | --- | | Overall Survival | | | | | Deaths (%) | 153 (63.0) | 153 (65.1) | 179 (73.1) | | Median (months) (95% CI) | 13.67 (11.10, 16.13) | 13.54 (11.24, 16.69) | 9.59 (8.77, 11.14) | | Hazard Ratio (95% CI)b | 0.69 (0.55, 0.86) | 0.73 (0.58, 0.90) | - | | p-value NT | 0.0007 | 0.0041 | - | | Progression-Free Survival | | | | | Disease Progression or Death (%) | 177 (72.8) | 180 (76.6) | 160 (65.3) | | Median (months) (95% CI) | 5.95 (5.49, 7.20) | 2.83 (2.17, 4.17) | 5.39 (4.17, 5.91) | | Hazard Ratio (95% CI)b | 0.79 (0.64, 0.98) | 1.21 (0.97, 1.50) | - | | p-value NT | 0.0353 | 0.0901 | - | | Overall Response Rate, n (%)a | 122 (50.2) | 64 (27.2) | 69 (28.2) | | (95% CI) | (43.7, 56.7) | (21.6, 33.4) | (22.6, 34.2) | | Complete Response (%) | 35 (14.4) | 30 (12.8) | 17 (6.9) | | Partial Response (%) | 87 (35.8) | 34 (14.5) | 52 (21.2) | | Duration of Response (months)a | | | | | Median (95% CI) | 8.18 (6.70, 11.37) | 12.65 (8.31, 27.43) | 7.13 (5.59, 8.48) | | Range | 1.5+, 35.9+ | 1.4+, 34.5+ | 1.4+, 31.8+ | a Assessed by BICR. b Based on stratified Cox proportional hazard model. Hazard ratios are reported for each OPDIVO containing arm compared to chemotherapy within each analysis population. NT: Not evaluated for statistical significance as per pre-specified hierarchical testing procedure. # c. Pediatric Extrapolation Patients aged 18 years and older were eligible for enrollment into the CHECKMATE-648 trial if they fulfilled the other inclusion criteria. The youngest patient enrolled in the study was 26 years old. For the CHECKMATE-648 study, data from adult patients aged 26 years an… --- **Source:** [https://fda-staging.innolitics.com/device/P150025S018](https://fda-staging.innolitics.com/device/P150025S018) **Published by [Innolitics](https://innolitics.com)** — a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices. 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