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Multiplex imaging of tumor tissue samples allows for improved biomarker discovery and increased accuracy in predicting patient therapeutic responses
Published 13th October, 2021
Following the success of monotherapy checkpoint inhibitors for cancer treatment, researchers are looking at combinations of established and new immunotherapies to extend the success of these treatments to increased patient numbers. However, leveraging the host immune system to fight cancer requires a more complete understanding of how components within the tumor microenvironment interact to promote or block tumor progression.
Previous methods used to analyze the tumor microenvironment, such as immunohistochemistry, can limit the number of biomarkers studied per sample. To overcome this limitation, many have transitioned to multiplex imaging. This application combines the study of multiple biomarkers in a tissue section with the capability to profile the spatial organization of each biomarker, either alone or in combination, to determine the tissue motifs of each tumor.
In this application note, we highlight work from Abcam's in-house research laboratories on multiplex IHC using a tyramide signal amplification(TSA) system and Drs Koei Chin and Joe Gray from the Department of Biomedical Engineering at the Oregon Health and Science University (OHSU), employing a cyclic immunofluorescence (CycIF) protocol, which demonstrates the successful implementation of Abcam antibodies to key immuno-oncology targets in different multiplex imaging solutions.
To determine the presence, localization, and abundance of six different immune checkpoints and immune cell phenotype markers in parallel, a selected panel of abcam antibodies in conjunction with Opal™ 7 color automation IHC kit from Akoya Biosciences (Table 1) was used.
Table 1. Antibody labeling and experimental conditions with PD-L1 / PD1 multiplex IHC-IF antibody panel (PD-L1, PD1, CD68, CD3, Ki67, panCK) (ab269812) used in multiplex staining. Antibodies within the panel are also available for individual purchase.
|Order of staining
|Recombinant anti-PD1 antibody (CAL20)
|Recombinant anti-PD-L1 antibody (CAL10)
|Recombinant anti-CD68 antibody (SP251)
|Anti-CD3 antibody (SP7)
|Recombinant anti-Ki67 antibody (SP6)
|Anti-pan cytokeratin antibody (C11)
To view the data and discover other high-quality reagents to support your multiplex experiments, download our application note below.