Spatially-resolved, Highly Multiplexed (up to 800-plex) Digital Characterization of Protein Distribution and Abundance in FFPE Tissue Sections Using Optical Barcoding
Joseph Beecham, PhD
Senior Vice President R&D, Nonostring Technologies
Intratumoral heterogeneity has emerged as a critical challenge to the implementation of targeted therapeutics. Historically, immunohistochemistry (IHC) has been used to assess spatial heterogeneity of proteins; however, it has been difficult to quantify protein abundance at high multiplex and wide dynamic range. Here, we report the development and validation of a spatially-resolved, antibody-based proteomic approach with a “barcoding-potential” to quantify up to 800 targets with 5.5 logs (base 10) of dynamic range in a single formalin-fixed paraffin-embedded (FFPE) slide.
By labeling antibodies with photocleavable oligos which are recognized by NanoString® nCounter® fluorescent barcodes and subsequently exposing them to focused UV light, we have developed an nCounter assay capable of quantifying protein abundance (and in future configurations, DNA and RNA targets) in a predefined spatial region of a tissue section.