A capture assay to analyse immunoglobulin secretion at the single-cell level

Area of Science:

• Immunology
• Cell Biology
• Biochemistry

Background:

• Existing secretion capture assays rely on paired monoclonal antibodies targeting cell surface antigens and secreted proteins.
• Measuring antibody secretion from B cells is crucial for understanding immune responses and antibody discovery.

Purpose of the Study:

• To develop a novel, accessible method for quantifying single-cell antibody secretion from B cells.
• To leverage the bivalent nature of antibodies for a new detection strategy.

Main Methods:

• Utilized an anti-immunoglobulin (Ig) antibody to bind membrane-bound Ig on B cells and capture secreted Ig.
• Employed a second, fluorophore-labeled anti-Ig antibody for detection of captured secreted Ig.
• Applied the assay to B cells undergoing differentiation into antibody-secreting plasma cells.

Main Results:

• Successfully demonstrated the capability of the assay to measure single-cell Ig secretion.
• Validated the method in a population of differentiating B cells.
• Showcased the assay's effectiveness in quantifying antibody secretion dynamics at the single-cell level.

Conclusions:

• The developed assay provides a reliable and accessible tool for single-cell antibody secretion analysis.
• This method has potential applications in immunological studies, antibody discovery, and functional profiling of B cell responses.