A multiplex implantable microdevice assay identifies synergistic combinations of cancer immunotherapies and conventional drugs
translate to 20%, 25%, 30% and 40% of drug concentration in PEG, respectively, when released from the nanowell. The calibration was determined previously using mass spectrometry measurements. Pure PEG was used in control conditions. Implanting multiple devices per tumor and/or multifocal animal models can increase the throughput up to 50–70 times as compared to conventional systemic treatment studies.
Before iterative cycles of staining, whole slide scanning and fluorophore bleaching, the slides were subjected to heat-mediated antigen retrieval by being immersed in citrate buffer for 25 minutes and then briefly rinsed in a hot bath and then immersed in Tris/EDTA buffer for 15 minutes, all using a Cuisinart Electric Pressure Cooker . Protein blocking was performed for 30 minutes at room temperature with 10% normal goat serum and 1% BSA in 1×PBS.
The readout antibody panel was carefully designed so that it broadly captures all major TME subtypes and allows to find synergy with the most established and/or emerging immunotherapies . Based on this, we defined a minimal essential set of 13 markers that classifies distinct myeloid and lymphoid lineages as well as components of non-immune stroma .
The cost of the MIMA workflow has two major components: one, the cost of the drug-loaded microdevices, which is ~$600–800 per device for a typical study, depending on the number and cost of individual drugs loaded into the device reservoirs; and two, the cost of the cycIF/mIHC, which is ~$50 per slide per cycle with basic immunohistochemistry infrastructure in place.
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