This study aims to optimize and validate a qPCR workflow for the reproducible and sensitive quantification of immune checkpoint transcripts (PD-1,PD-L1, CTLA-4) in FFPE lung cancer tissues. By refining tissue preparation, RNA input, and assay conditions, we establish a robust method for detecting gene expression across a broad dynamic range, enabling reliable assessment of immunotherapy response and supporting biomarker-driven patient stratification.
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