Multiplex immunohistochemistry (IHC) techniques allow multiple proteins to be visualized in the tissue leading to the identification of cell phenotypes, which can help gain an understanding of the spatial arrangement of cells within a tumor. Each multiplex IHC method places unique challenges upon antibody performance. For example, tyramide signal amplification (TSA)-based multiplex immunofluorescence assays (mIF) require optimization of staining order, while antibody performance can become compromised after metal conjugation during imaging mass spectrometry.
In this on-demand SelectScience® webinar, Mike Spencer, senior director of antibody validation and IHC at Fortis Life Sciences, discusses effective strategies to ensure optimal antibody performance in multiplex IHC assays.
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MS: Yes, minimally one should be able to replicate the data generated by the vendor. In addition, one needs to determine if the antibody is fit for purpose within their assay. This includes confirming that the antibody works with the reagents and samples being used in the assay.
MS: Not in all cases. Each multiplex system has specific restraints that may alter antibody activity. A good example is techniques that use conjugated antibodies. Conjugation may impact the ability of the antibody to bind to its target.
MS: They are not pre-conjugated.
MS: In most cases antibodies that have been validated using citrate retrieval will work just fine with Tris retrieval. In fact, you may get a boost in signal. One should monitor background staining which can also be increased with the use of Tris.
MS: Currently, multiplex techniques are not approved for clinical diagnostic use. Multiplex is being used to drive disease research forward in the basic science and translational science field.
MS: Multiplex assays can be used to profile patients to understand disease progression and predict what therapies they might respond to.
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