Uncover antibody-secreting cells based on antigen-binding specificity using fluorescent detection and Cyto-Mine® Chroma.
Author: Benjamin Duckless
Senior Field Application Scientist, Sphere Bio
Keywords: Cell line development, antibody discovery
Introduction
As antibody discovery and cell line development landscapes increase in complexity, researchers require more reliable tools at their disposal to aid in faster, more streamlined decisions. Sphere Fluidics has answered this call with the newly launched Cyto-Mine® Chroma. Outfitted with enhanced detection parameters, a new user interface and the tried-and-true microfluidic picodroplet technology; the Cyto-Mine® Chroma enables the exploration of complex biological questions across millions of cells in a refined and approachable workflow.
This study presented showcases how Cyto-Mine® Chroma can be utilized to run functional bioassays requiring multiplexing to identify picodroplets containing antibody secreting cells, target cells and the secreted antibody bound to the surface receptor on the target cells.
Key Findings
ANTIGEN-SPECIFIC MULTIPLEX ASSAYS
∙ Blue Fluorescence (eFL450)- Identifies antibody-secreting hybridoma cells
∙ Far Red Fluorescence (eFL670) – Identifies the target cells
∙ Green Fluorescence (AF488) – Detection antibody to identify binding of the secreted antibody with the corresponding receptors on the target cells
CO-ENCAPSULATION TECHNOLOGY enables functional assays by creating millions of picolitre sized reaction vessels in a single experiment that contain both antibody secreting cells and target cells.
OFF-LINE FLUORESCENT IMAGING confirms the presence of all three key components of a sorted picodroplet that meets the selection criteria and has been dispensed from the Cyto-Mine® Chroma to a microplate.
Study Insights
By expanding fluorescence detection capabilities to four channels and introducing serial gating during the sorting procedure, Cyto-Mine® Chroma provides a more refined approach to screening and selecting antibody secreting cells of interest. The utilization of an antigen specific binding assay takes it one level further by not only selecting cells that are producing antibodies, but antibodies that actively bind to their receptor on target cells that have been co-encapsulated with the antibody secreting cells within the picodroplets.
Conclusion
Cyto-Mine® Chroma represents a major leap forward in single-cell analysis. This study showcases its ability to improve assay resolution, increase screening efficiency, and support next-generation therapeutic development.
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The Next Generation: Cyto-Mine® Chroma
Automate. Accelerate. Analyze millions of cells in a single day.
Think Cyto-Mine®, but supercharged, enabling multiplexing and greater assay flexibility to fit your needs. It means that you can examine vastly greater numbers of cells — and isolate the most valuable ones— with unparalleled precision.
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