A recent study comparing immunooncology biomarker types has demonstrated that multiplex immunofluorescence with spatial characterization significantly outperforms other biomarker testing methods. The meta-analysis performed by scientists at Johns Hopkins University, Yale University, and other institutions determined that multiplex immunofluorescence is better than gene expression profiling, tumor mutational burden assessment, and immunohistochemistry for predicting patient response to treatments targeting PD-1/PD-L1.1

A new type of biomarker assay, multiplex immunofluorescence allows investigators to simultaneously analyze the expression of many proteins in individual cells within the tumor microenvironment, preserving critical information about which cells are active and how they are spatially distributed relative to one another. This type of analysis is made possible with the end-to-end solutions offered by the Codex ultrahigh multiplexing platform for biomarker discovery, and the Phenoptics high-throughput multiplexing platform for translational and clinical research, both from Akoya Biosciences, Hopkinton, Mass.

Immunotherapies targeting PD-1 or PD-L1 have proven effective for treating cancer in some patients, but there remains a paucity of accurate biomarkers that can differentiate responders from nonresponders. Identifying the patients most likely to respond to these therapies is an important step in ensuring optimal outcomes for all patients. To date, several assays have been developed with the potential to predict response based on genetic signatures, gene expression, or immunohistochemistry. Although these assays are helpful in limited situations, there is a need for options that are better at predicting response across a larger percentage of cases.


Cliff Hoyt, Akoya Biosciences.

The collaborative report, which also involved scientists at Vanderbilt and Northwestern universities, reviewed published data from more than 50 studies covering more than 10 types of cancer and over 8,000 patients. Statistical analyses were performed to assess the performance and predictive value of each type of biomarker. Tumor mutational burden, gene expression profiling, and immunohistochemistry all performed comparably in differentiating between responders and nonresponders, while multiplex immunofluorescence had considerably better performance metrics than these three assay types. Specifically, it had fewer false positives, meaning it was less likely to predict positive response in a patient who would not ultimately respond to therapy.

“This meta-analysis of previous studies clearly demonstrates the potential for using multiplex immunofluorescence to generate more comprehensive and reliable data to better predict response to anti-PD-1/PD-L1 treatments,” says Cliff Hoyt, vice president of translational and scientific affairs at Akoya and a coauthor of the paper. “This contributes to growing evidence that spatial resolution of tumor biomarkers is essential for an accurate view of cancer biology, and the Akoya team is excited to help researchers continue down this promising avenue of investigation.”

By performing an sROC curve evaluation, the study’s authors cited multiplex immunofluorescence and multiplex immunohistochemistry (AUC of 0.79) as having significantly higher diagnostic predictive accuracy when compared with PD-L1 immunohistochemistry (AUC of 0.65, PAkoya Biosciences.


  1. Lu S, Stein JE, Rimm DL, et al. Comparison of biomarker modalities for predicting response to PD-1/PD-L1 checkpoint blockade: a systematic review and meta-analysis. JAMA Oncol. Epub ahead of print, July 18, 2019; doi: 10.1001/jamaoncol.2019.1549.

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A member of the Phenoptics portfolio acquired by Akoya Biosciences from PerkinElmer, the Vectra Polaris system provides the high throughput and standardization required to support large-scale translational studies.