A Workflow for Multiplex Protein Imaging with Co-detection of Low Abundant Molecules and Proteins

Value Proposition

• Multiplex Capability: Enables iterative TSA applications, a capability previously unattainable in highly multiplexed platforms which are currently capable of only eight markers per section.
• High Sensitivity: Detects lowly-abundant proteins to map the full complexity of tissues architecture without degrading tissue morphology or antigenicity between staining cycles.
• Compatibility with traditional spatial proteomics workflows: Integrates TSA-based signal amplification into high-dimensional spatial proteomics workflows such as IMC, CyCIF, & CODEX.
• Broadly Applicable: can be expanded to study a wide range of biological processes in situ.

Unmet Need

• While highly multiplexed immuno-imaging is critical for delineating tissue architecture and disease pathology, current platforms lack the sensitivity to detect low-abundance proteins essential for unraveling complex disease mechanisms.
• Detecting low abundance proteins with sufficient sensitivity remains a major challenge for high-plex imaging platforms, limiting insights into key disease pathways within tissue environments.
• Existing platforms either prioritize broad multiplexing or sensitive detection of low abundance proteins, making it difficult to simultaneously visualize rare biomarkers within their complex tissue environments.
• This highlights an urgent need for next-generation imaging technologies that combine high sensitivity for low-abundance proteins with robust multiplexing to preserve spatial and molecular context.

Technology Description

• Researchers at Johns Hopkins have developed a workflow that achieves both high sensitivity and high multiplexed detection within the same section of formalin-fixed, paraffin-embedded (FFPE) tissues.
• The disclosed technology uses optimized bleaching technology to enable iterative TSA applications, without degrading tissue morphology or antigenicity, a capability previously unattainable in highly multiplexed platforms.
• Researchers at Johns Hopkins have demonstrated six-plex detection of low abundant senescence markers in archival FFPE tissues, and integration of the disclosed technology with traditional high-plex techniques (CyCIF, CODEX, and IMC) was able to achieve 40+ plex detections while preserving sensitivity.

Stage of Development

• The disclosed technology has reached a stage where the workflow is fully established

Data Availability: Data available upon request.

Publication: N/a