Development of a Comprehensive Antibody Staining Database Using a Standardized Analytics Pipeline.

Area of Science:

• Immunology
• Biotechnology
• Computational Biology

Background:

• Large-scale immune monitoring is crucial for biomarker discovery and patient stratification in immunotherapy.
• Mass cytometry is a key technology for immune monitoring, but requires standardized workflows for large-scale applications.

Purpose of the Study:

• To establish a standardized workflow for the acquisition and analysis of large-scale mass cytometry experiments.
• To create a reproducible and scalable method for immune monitoring in clinical trials.

Main Methods:

• Implementation of a two-tiered barcoding system.
• Utilization of a broad lyophilized antibody panel.
• Integration of a fully automated, cloud-based analysis platform.
• Application of the workflow to a large antibody staining screen (LEGENDScreen kit).

Main Results:

• Generated single-cell data for 350 antibodies across 71 profiling subsets.
• Validated known immune system trends and identified potential markers for MAIT cells.
• Assessed the impact of fixation on staining intensity, noting signal gains or losses for specific markers.
• Demonstrated the workflow's applicability to large antibody staining screens.

Conclusions:

• The proposed standardized workflow is suitable for large-scale mass cytometry experiments and can be integrated into clinical trials.
• The workflow facilitates biomarker discovery and responder stratification in immunotherapy.
• The generated antibody staining dataset serves as a valuable online resource for researchers.